Anti tubercular drug: compostions and methods

ABSTRACT

Methods and compositions for treating disease caused by infectious agents, particularly tuberculosis. In particular, methods and compositions comprising substituted ethylene diamines for the treatment of infectious diseases are provided. In one embodiment, these methods and compositions are used for the treatment of mycobacterial infections, including, but not limited to, tuberculosis.

CROSS REFERENCE TO RELATED PATENT APPLICATIONS

[0001] The present application is a continuation-in-part application ofpending U.S. patent application Ser. No. 10/147,587 filed May 17, 2002.The present application also claims priority to U.S. Provisional PatentApplication Serial No. 60/381,220 filed May 17, 2002.

FIELD OF INVENTION

[0002] The present invention relates to methods and compositions fortreating disease caused by microorganisms, particularly tuberculosis.The present invention also relates to methods and compositions havingimproved anti-mycobacterial activity, namely compositions comprisingnovel substituted ethylene diamine compounds.

BACKGROUND OF THE INVENTION

[0003] Mycobacterial infections often manifest as diseases such astuberculosis. Human infections caused by mycobacteria have beenwidespread since ancient times, and tuberculosis remains a leading causeof death today. Although the incidence of the disease declined, inparallel with advancing standards of living, since the mid-nineteenthcentury, mycobacterial diseases still constitute a leading cause ofmorbidity and mortality in countries with limited medical resources.Additionally, mycobacterial diseases can cause overwhelming,disseminated disease in immunocompromised patients. In spite of theefforts of numerous health organizations worldwide, the eradication ofmycobacterial diseases has never been achieved, nor is eradicationimminent. Nearly one third of the world's population is infected withmycobacterium tuberculosis complex, commonly referred to as tuberculosis(TB), with approximately 8 million new cases, and two to three milliondeaths attributable to TB yearly. Tuberculosis (TB) is the cause of thelargest number of human deaths attributable to a single etiologic agent(see Dye et al., J. Am. Med. Association, 282, 677-686, (1999); and 2000WHO/OMS Press Release).

[0004] After decades of decline, TB is now on the rise. In the UnitedStates, up to 10 million individuals are believed to be infected. Almost28,000 new cases were reported in 1990, constituting a 9.4 percentincrease over 1989. A sixteen percent increase in TB cases was observedfrom 1985 to 1990. Overcrowded living conditions and shared air spacesare especially conducive to the spread of TB, contributing to theincrease in instances that have been observed among prison inmates, andamong the homeless in larger U.S. cities. Approximately half of allpatients with “Acquired Immune Deficiency Syndrome” (AIDS) will acquirea mycobacterial infection, with TB being an especially devastatingcomplication. AIDS patients are at higher risks of developing clinicalTB, and anti-TB treatment seems to be less effective than in non-AIDSpatients. Consequently, the infection often progresses to a fataldisseminated disease.

[0005] Mycobacteria other than M. tuberculosis are increasingly found inopportunistic infections that plague the AIDS patient. Organisms fromthe M. avium-intracellulare complex (MAC), especially serotypes four andeight, account for 68% of the mycobacterial isolates from AIDS patients.Enormous numbers of MAC are found (up to 10¹⁰ acid-fast bacilli per gramof tissue), and consequently, the prognosis for the infected AIDSpatient is poor.

[0006] The World Health Organization (WHO) continues to encourage thebattle against TB, recommending prevention initiatives such as the“Expanded Program on Immunization” (EPI), and therapeutic complianceinitiatives such as “Directly Observed Treatment Short-Course” (DOTS).For the eradication of TB, diagnosis, treatment, and prevention areequally important. Rapid detection of active TB patients will lead toearly treatment by which about 90% cure is expected. Therefore, earlydiagnosis is critical for the battle against TB. In addition,therapeutic compliance will ensure not only elimination of infection,but also reduction in the emergence of drug-resistance strains.

[0007] The emergence of drug-resistant M. tuberculosis is an extremelydisturbing phenomenon. The rate of new TB cases proven resistant to atleast one standard drug increased from 10 percent in the early 1980's to23 percent in 1991. Compliance with therapeutic regimens, therefore, isalso a crucial component in efforts to eliminate TB and prevent theemergence of drug resistant strains. Equally important is thedevelopment of new therapeutic agents that are effective as vaccines,and as treatments, for disease caused by drug resistant strains ofmycobacteria.

[0008] Although over 37 species of mycobacteria have been identified,more than 95% of all human infections are caused by six species ofmycobacteria: M. tuberculosis, M. avium intracellulare, M. kansasii, M.fortuitum, M. chelonae, and M. leprae. The most prevalent mycobacterialdisease in humans is tuberculosis (TB) which is predominantly caused bymycobacterial species comprising M. tuberculosis, M. bovis, or M.africanum (Merck Manual 1992). Infection is typically initiated by theinhalation of infectious particles which are able to reach the terminalpathways in lungs. Following engulfment by alveolar macrophages, thebacilli are able to replicate freely, with eventual destruction of thephagocytic cells. A cascade effect ensues wherein destruction of thephagocytic cells causes additional macrophages and lymphocytes tomigrate to the site of infection, where they too are ultimatelyeliminated. The disease is further disseminated during the initialstages by the infected macrophages which travel to local lymph nodes, aswell as into the blood stream and other tissues such as the bone marrow,spleen, kidneys, bone and central nervous system. (See Murray et al.Medical Microbiology, The C.V. Mosby Company 219-230 (1990)).

[0009] There is still no clear understanding of the factors whichcontribute to the virulence of mycobacteria. Many investigators haveimplicated lipids of the cell wall and bacterial surface as contributorsto colony morphology and virulence. Evidence suggests that C-mycosides,on the surface of certain mycobacterial cells, are important infacilitating survival of the organism within macrophages. Trehalose 6,6′dimycolate, a cord factor, has been implicated for other mycobacteria.

[0010] The interrelationship of colony morphology and virulence isparticularly pronounced in M. avium. M. avium bacilli occur in severaldistinct colony forms. Bacilli which grow as transparent, or rough,colonies on conventional laboratory media are multiplicable withinmacrophages in tissue culture, are virulent when injected intosusceptible mice, and are resistant to antibiotics. Rough or transparentbacilli, which are maintained on laboratory culture media, oftenspontaneously assume an opaque R colony morphology, at which time theyare not multiplicable in macrophages, are avirulent in mice, and arehighly susceptible to antibiotics. The differences in colony morphologybetween the transparent, rough and opaque strains of M. avium are almostcertainly due to the presence of a glycolipid coating on the surface oftransparent and rough organisms which acts as a protective capsule. Thiscapsule, or coating, is composed primarily of C-mycosides whichapparently shield the virulent M. avium organisms from lysosomal enzymesand antibiotics. By contrast, the non-virulent opaque forms of M. aviumhave very little C-mycoside on their surface. Both the resistance toantibiotics and the resistance to killing by macrophages have beenattributed to the glycolipid barrier on the surface of M. avium.

[0011] Diagnosis of mycobacterial infection is confirmed by theisolation and identification of the pathogen, although conventionaldiagnosis is based on sputum smears, chest X-ray examination (CXR), andclinical symptoms. Isolation of mycobacteria on a medium takes as longas four to eight weeks. Species identification takes a further twoweeks. There are several other techniques for detecting mycobacteriasuch as the polymerase chain reaction (PCR), mycobacterium tuberculosisdirect test, or amplified mycobacterium tuberculosis direct test (MTD),and detection assays that utilize radioactive labels.

[0012] One diagnostic test that is widely used for detecting infectionscaused by M. tuberculosis is the tuberculin skin test. Although numerousversions of the skin test are available, typically one of twopreparations of tuberculin antigens are used: old tuberculin (OT), orpurified protein derivative (PPD). The antigen preparation is eitherinjected into the skin intradermally, or is topically applied and isthen invasively transported into the skin with the use of a multipronginoculator (Tine test). Several problems exist with the skin testdiagnosis method. For example, the Tine test is not generallyrecommended because the amount of antigen injected into the intradermallayer cannot be accurately controlled. (See Murray et al. MedicalMicrobiology, The C.V. Mosby Company 219-230 (1990)).

[0013] Although the tuberculin skin tests are widely used, theytypically require two to three days to generate results, and many times,the results are inaccurate since false positives are sometimes seen insubjects who have been exposed to mycobacteria, but are healthy. Inaddition, instances of mis-diagnosis are frequent since a positiveresult is observed not only in active TB patients, but also in personsvaccinated with Bacille Calmette-Guerin (BCG), and those who had beeninfected with mycobacteria, but have not developed the disease. It ishard therefore, to distinguish active TB patients from the others, suchas household TB contacts, by the tuberculin skin test. Additionally, thetuberculin test often produces a cross-reaction in those individuals whowere infected with mycobacteria other than M. tuberculosis (MOTT).Therefore, diagnosis using the skin tests currently available isfrequently subject to error and inaccuracies.

[0014] The standard treatment for tuberculosis caused by drug-sensitiveorganisms is a six-month regimen consisting of four drugs given for twomonths, followed by two drugs given for four months. The two mostimportant drugs, given throughout the six-month course of therapy, areisoniazid and rifampin. Although the regimen is relatively simple, itsadministration is quite complicated. Daily ingestion of eight or ninepills is often required during the first phase of therapy; a dauntingand confusing prospect. Even severely ill patients are often symptomfree within a few weeks, and nearly all appear to be cured within a fewmonths. If the treatment is not continued to completion, however, thepatient may experience a relapse, and the relapse rate for patients whodo not continue treatment to completion is high. A variety of forms ofpatient-centered care are used to promote adherence with therapy. Themost effective way of ensuring that patients are taking their medicationis to use directly observed therapy, which involves having a member ofthe health care team observe the patient take each dose of each drug.Directly observed therapy can be provided in the clinic, the patient'sresidence, or any mutually agreed upon site. Nearly all patients whohave tuberculosis caused by drug-sensitive organisms, and who completetherapy will be cured, and the risk of relapse is very low (“EndingNeglect: The Elimination of Tuberculosis in the United States” ed. L.Geiter Committee on the Elimination of Tuberculosis in the United StatesDivision of Health Promotion and Disease Prevention, Institute ofMedicine. Unpublished.)

[0015] What is needed are effective therapeutic regimens that includeimproved vaccination and treatment protocols. Currently availabletherapeutics are no longer consistently effective as a result of theproblems with treatment compliance, and these compliance problemscontribute to the development of drug resistant mycobacterial strains.

[0016] Ethambutol (EMB) is a widely used antibiotic for the treatment ofTB, with over 300 million doses delivered for tuberculosis therapy in1988.

[0017] Ethambutol, developed by Lederle Laboratories in the 1950s, haslow toxicity and is a good pharmacokinetic. However, ethambutol has arelatively high Minimum Inhibition Concentration (MIC) of about 5 μg/ml,and can cause optic neuritis. Thus, there is an increasing need for new,and more effective, therapeutic compositions (See for example, U.S. Pat.No. 3,176,040, U.S. Pat. No. 4,262,122; U.S. Pat. No. 4,006,234; U.S.Pat. No. 3,931,157; U.S. Pat. No. 3,931,152; U.S. Re. Pat. No. 29,358;and Häusler et al., Bioorganic & Medicinal Chemistry Letters 11 (2001)1679-1681). In the decoder years since the discovery of the beneficialeffects of ethambutol, few pharmacological advances in TB treatment havebeen developed. Moreover, with the combined emergence of drug resistantstrains, and the more prevalent spread of mycobacterial disease, it isbecoming seriously apparent that new therapeutic compositions arecrucial in the fight against tuberculosis.

[0018] Clearly effective therapeutic regimens that include improvedvaccination and treatment protocols are needed. A therapeutic vaccinethat would prevent the onset of tuberculosis, and therefore eliminatethe need for therapy is desirable. Although currently availabletherapeutics such as ethambutol are effective, the emergence of drugresistant strains has necessitated new formulations and compositionsthat are more versatile than ethambutol. Currently availabletherapeutics are no longer consistently effective as a result of theproblems with treatment compliance, lending to the development of drugresistant mycobacterial strains. What is needed are new anti-tuberculardrugs that provide highly effective treatment, and shorten or simplifytuberculosis chemotherapy.

SUMMARY OF THE INVENTION

[0019] The present invention comprises methods and compositionscomprising ethylene diamine compounds effective for the treatment ofinfectious disease. The present invention also provides methods andcompositions comprising substituted ethylene diamines having improvedanti-mycobacterial activity, including substituted ethylene diamineshaving improved anti-tuberculosis activity.

[0020] The present invention contemplates substituted ethylene diamines,which can derive from a variety of amine compounds. In the presentinvention, the substituted ethylene diamines are based on the followingstructure.

[0021] The substituted ethylene diamine compounds described herein aresynthesized and screened for activity as follows. A chemical library ofsubstituted ethylene diamines is prepared on a solid polystyrene supportusing split and pool technologies. This technique allows for thesynthesis of a diverse set of substituted ethylene diamines. Thesediamines are screened for anti-TB activity using in vitro, biologicalassays, including a High-Throughput Screening (HTS) assay, based on therecently completed genomic sequence of M. tuberculosis, and a MinimumInhibition Concentration (MIC) assay.

[0022] The methods and compositions described herein comprisesubstituted ethylene diamines that are effective against disease causedby infectious organisms, including, but not limited to, bacteria andviruses. One embodiment of the invention provides methods andcompositions comprising substituted ethylene diamines that are effectiveagainst mycobacterial disease. Another embodiment of the inventionprovides methods and compositions comprising substituted ethylenediamines that have MIC of 50 μM or lower for mycobacterial disease.Another embodiment of the present invention comprises substitutedethylene diamines that have an MIC of 25 μM or lower for mycobacterialdisease. Yet another embodiment of the present invention comprisessubstituted ethylene diamines that have an MIC of 12.5 μM or lower formycobacterial disease. Another embodiment of the present inventioncomprises substituted ethylene diamines that have an MIC of 5 μM orlower for mycobacterial disease In another embodiment of the presentinvention, the methods and compositions comprise substituted ethylenediamines with HTS Luc activity of 10% or greater. In yet anotherembodiment of the present invention, the methods and compositionscomprise substituted ethylene diamines, wherein one amine group isderived from a primary amine, and wherein the other amine group isderived from a primary or secondary amine. In another embodiment of thepresent invention, the methods and compositions comprise substitutedethylene diamines, wherein one amine is derived fromcis-(−)myrtanylamine, cyclooctylamine, 2,2-diphenylethylamine,3,3-diphenylpropylamine, (+)-bomylamine, 1-adamantanemethylamine,(+)-isopinocampheylamine; or (−)-isopinocampheylamine.

[0023] The present invention contemplates various salt complexes andother substituted derivatives of the substituted ethylene diamines. Thepresent invention also contemplates enantiomers and other stereoisomersof the substituted ethylene diamines and their substituted derivatives.The present invention further contemplates treatment for animals,including, but not limited to, humans.

[0024] Accordingly, it is an object of the present invention to providemethods and compositions for the treatment and prevention of diseasescaused by microorganisms.

[0025] Accordingly, it is an object of the present invention to providemethods and compositions for the treatment and prevention of infectiousdiseases.

[0026] Another object of the present invention is to provide methods andcompositions for the treatment and prevention of mycobacterial disease,including but not limited to, tuberculosis.

[0027] Yet another object of the present invention is to provide methodsand compositions for the treatment and prevention of infectious diseasesusing compositions comprising substituted ethylene diamines.

[0028] Another object of the present invention is to provide methods andcompositions for the treatment and prevention of mycobacterial diseaseusing compositions comprising substituted ethylene diamines.

[0029] Still another object of the present invention is to providemethods and compositions for the treatment and prevention oftuberculosis using compositions comprising substituted ethylenediamines.

[0030] Another object of the present invention is to provide methods andcompositions for the treatment and prevention of tuberculosis usingcompositions comprising substituted ethylene diamines, wherein thediamine has an MIC of 50 μM, or less.

[0031] Another object of the present invention is to provide methods andcompositions for the treatment and prevention of tuberculosis usingcompositions comprising substituted ethylene diamines, wherein thediamine has an MIC of 25 μM, or less.

[0032] Another object of the present invention is to provide methods andcompositions for the treatment and prevention of tuberculosis usingcompositions comprising substituted ethylene diamines, wherein thediamine has an MIC of 12.5 μM, or less.

[0033] Yet another object of the present invention is to provide methodsand compositions for the treatment and prevention of tuberculosis usingcompositions comprising substituted ethylene diamines, wherein thediamine has an MIC of 5 μM, or less.

[0034] Yet another object of the present invention is to provide methodsand compositions for the treatment and prevention of tuberculosis usingcompositions comprising substituted ethylene diamines, wherein thediamine has HTS/Luc activity of 10% or greater.

[0035] Another object of the present invention is to provide methods andcompositions for the treatment and prevention of tuberculosis usingcompositions comprising substituted ethylene diamines, wherein one aminegroup is derived from a primary amine, and the other amine group isderived from a primary or secondary amine.

[0036] Yet another object of the present invention is to provide methodsand compositions for the treatment and/or prevention of tuberculosisusing compositions comprising substituted ethylene diamines, wherein oneamine is derived from cis-(−)myrtanylamine, cyclooctylamine,2,2-diphenylethylamine, 3,3-diphenylpropylamine, (+)-bomylamine,1-adamantanemethylamine, (+)-isopinocampheylamine; or(−)-isopinocampheylamine.

[0037] Yet another object of the present invention is to providecomposition for the therapeutic formulation for the treatment andprevention of mycobacterial disease.

[0038] Another object of the present invention is to providecompositions for therapeutic formulations for the treatment andprevention of mycobacterial disease caused by mycobacterial speciescomprising M. tuberculosis complex, M. avium intracellulare, M.kansarii, M. fortuitum, M. chelonoe, M. leprae, M. africanum, M.microti, or M. bovis.

[0039] These and other objects, features and advantages of the presentinvention will become apparent after a review of the following detaileddescription of the disclosed embodiments and the appended claims.

BRIEF DESCRIPTION OF THE FIGURES

[0040]FIG. 1 represents a flow chart schematic showing various solidsupport syntheses used to prepare substituted ethylene diamines.

[0041] FIGS. 2(a)-2(ac) provide chemical structures of a variety ofprimary amines.

[0042] FIGS. 3(a)-3(f) provide chemical structures of a variety ofacyclic secondary amines.

[0043] FIGS. 4(a)-4(i) provide chemical structures of a variety ofcyclic secondary amines.

[0044]FIG. 5 represents a flow schematic for a representative reactionpool of ten substituted ethylene diamines.

[0045]FIG. 6 is a graph of Luminescence Count per Second (LCPS) versusconcentration showing HTS Luc assay results for pooled substitutedethylene diamine compounds.

[0046]FIG. 7 is a graph of LCPS versus concentration showing HTS Lucassay results for individual substituted ethylene diamine compounds.

[0047]FIG. 8 is a graph of LCPS versus concentration showing HTS Lucassay results for individual substituted ethylene diamine compounds.

[0048]FIG. 9 is a bar graph providing a summary of MIC activities fordiscrete substituted ethylene diamines.

[0049]FIG. 10 is a bar graph providing a summary of Luciferase activityof discrete substituted ethylene diamines with at least 10% activity inreference to ethambutol at 3.1 μM.

[0050]FIG. 11 is a bar graph showing the frequency of occurrences of theselected amine monomers in the substituted ethylene diamine compoundsthat were active against TB. Amine monomers are represented by theirnumerical designations.

[0051]FIG. 12 represents a flow schematic showing a synthesis ofN-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine (compound 109).

[0052]FIG. 13 is a flow schematic showing a synthesis ofN-(Cyclooctyl)-N′-(1R, 2R, 3R, 5S)-(−)-isopinocampheylethane-1,2-diamineas hydrochloride (compound 59).

[0053]FIG. 14 is a mass spec profile for one representative sample wellcontaining pooled substituted ethylene diamine compounds.

[0054]FIG. 15 is a mass spec profile for compound 109,N-Geranyl-N′-(2-adamanthyl) ethane-1,2-diamine.

[0055]FIG. 16 is a proton NMR profile for compound 109,N-Geranyl-N′-(2-adamanthyl) ethane-1,2-diamine.

[0056]FIG. 17 is a bar graph of data from a Colony Forming Units/Lung(CFU/Lung) study showing CFU/Lung growth over time in days for variouscompounds.

[0057]FIG. 18 is a bar graph of data from a CFU/Lung study showingCFU/Lung growth over time in days for various compounds.

[0058]FIG. 19 is a bar graph of data from a CFU/Lung study showingCFU/Lung growth over time in days for various compounds.

[0059]FIG. 20 is a bar graph of data from a lesion study showing visiblelesions over time after treatment with various compounds.

[0060]FIG. 21 provides a schematic demonstrating the identification of adrug candidate

[0061]FIG. 22 provides the compounds tested for in vivo efficacy.

[0062]FIG. 23 is a graph showing the results of in vivo studies ofcompounds 73 and 109 at 1 and 10 mg/kg doses (spleen).

[0063]FIG. 24 is a graph showing the results of in vivo studies ofcompounds 73 and 109 at 1 and 10 mg/kg doses (lungs).

[0064]FIG. 25 is a graph showing in vivo studies of compounds 59 and 111at 1 and 10 mg/kg doses (spleen).

[0065]FIG. 26 is a graph showing in vivo studies of compounds 59 and 111at 1 and 10 mg/kg doses (lungs).

[0066]FIG. 27 is a graph showing the results of efficacy testing of thecompounds 58, 73, 109, and 111 in C57BL.6 mice infected with M.tuberculosis H37Rv (spleen). Mice were infected i.v. with 5×10⁶ CFU M.tuberculosis H37Rv; treatment with drugs started 18 days followinginfection. EC-EC—early control, CFU in lungs of mice at the day ofchemotherapy start. Mice received: 1—untreated mice, 2—INH (25 mg/kg),3—EMB (100 mg/kg), 4—comp. 109 (25 mg/kg), 4*—comp.109 (10 mg/kg),4**—comp. 109 (0.1 mg/kg), 5—comp. 58 (25 mg/kg), 6—comp.73 (25 mg/kg),7—comp. 111 (25 mg/kg).

[0067]FIG. 28 is a graph showing the results of efficacy testing of thecompounds 58, 73, 109, and 111 in C57BL.6 mice infected with M.tuberculosis H37Rv (lungs). Mice were infected i.v. with 5×10⁶ CFU M.tuberculosis H37Rv; treatment with drugs started 18 days followinginfection. EC-EC—early control, CFU in lungs of mice at the day ofchemotherapy start. Mice received: 1—untreated mice, 2—INH (25 mg/kg),3—EMB (100 mg/kg), 4—comp. 109 (25 mg/kg), 4*—comp.109 (10 mg/kg),4**—comp. 109 (0.1 mg/kg), 5—comp. 58 (25 mg/kg), 6—comp.73 (25 mg/kg),7—comp. 111 (25 mg/kg).

[0068]FIG. 29 provides LC/MS data of tested compounds.

[0069]FIG. 30 provides a graph showing results of PK studies with acassette dosing of tested compounds to mice. Oral delivery. Compound NSC722039 in the study reads as the compound 37, NSC 722040—compound 59,NSC 722041—compound 109.

[0070]FIG. 31 provides a graph showing results of PK studies with acassette dosing of tested compounds to mice. Peritoneal delivery.Compound NSC 722039 in the study reads as the compound 37, NSC722040—compound 59, NSC 722041—compound 109.

[0071]FIG. 32 provides a graph showing results of PK studies with acassette dosing of tested compounds to mice. Intravenous delivery.Compound NSC 722039 in the study reads as the compound 37, NSC722040—compound 59, NSC 722041—compound 109.

[0072]FIG. 33 provides a graph showing the results of PK Studies of thecompound 109 in mice.

[0073]FIG. 34. Tissue distribution of 109 in mice (i.v., 3 mg/kg).

[0074]FIG. 35. Tissue distribution of 109 in mice (p.o., 25 mg/kg).

[0075]FIG. 36 Metabolism of the compound 109 in mouse urine.

[0076]FIG. 37. No glucoronidation metabolites of 109 were found in mouseurine.

[0077]FIG. 41. Scheme 1. Synthesis of 100,000 compound library ofethambutol analogues on solid support.

[0078]FIG. 41. Scheme 2. Attempts to synthesize SQBisAd on solidsupport.

[0079]FIG. 42 provides structures of representative targeted diaminesprepared via acylation by amino acids.

[0080]FIG. 43 provides Table 25 summarizing data for synthesized platesof diamines for the prepared library of targeted 20,000 ethambutolanalogs.

[0081]FIG. 44 provides Scheme 5 showing the synthesis of the diaminelibrary using amino acids as linkers.

[0082]FIG. 45 provides a schematic showing the occurrence of aminemonomers in the hits that were generated in the original 100,000compound library of EMB analogs.

[0083]FIG. 46 provides a schematic showing structural diversity amongprimary amines.

[0084]FIG. 47 provides Table 26 listing the amino acids that were usedin the prepartion of the diamine library.

[0085]FIG. 48 provides carbonyl compounds used as reagents in thesynthesis of the diamine library.

[0086]FIG. 49 provides Table 27 showing carbonyl compounds used in themasterplate for the synthesis of the diamine library.

[0087]FIG. 50 provides representative examples of MIC and Lux data forthe diamine library.

[0088]FIG. 51 provides a schematic showing the occurrence of alkylatingmonomers in final diamine products with anti-TB activity.

[0089]FIG. 52 provides the layout of a representative 96-welldeconvolution plate.

[0090]FIG. 53 provides a list of compound hits and structures for themodified linker diamine library.

DETAILED DESCRIPTION

[0091] The present invention may be understood more readily by referenceto the following detailed description of the specific embodimentsincluded herein. However, although the present invention has beendescribed with reference to specific details of certain embodimentsthereof, it is not intended that such details should be regarded aslimitations upon the scope of the invention. The entire text of thereferences mentioned herein are hereby incorporated in their entiretiesby reference including U.S. patent application Ser. No. 10/147,587 filedMay 17, 2002, and U.S. Provisional Patent Application Serial No.60/381,220 filed May 17, 2002.

[0092] Mycobacterial infections, such as those causing tuberculosis,once thought to be declining in occurrence, have rebounded, and againconstitute a serious health threat. Tuberculosis (TB) is the cause ofthe largest number of human deaths attributed to a single etiologicagent with two to three million people infected with tuberculosis dyingeach year. Areas where humans are crowded together, or living insubstandard housing, are increasingly found to have persons affectedwith mycobacteria. Individuals who are immunocompromised are at greatrisk of being infected with mycobacteria and dying from such infection.In addition, the emergence of drug-resistant strains of mycobacteria hasled to treatment problems of such infected persons.

[0093] Many people who are infected with mycobacteria are poor, or livein areas with inadequate healthcare facilities. As a result of variousobstacles (economical, education levels, etc.), many of theseindividuals are unable to comply with the prescribed therapeuticregimens. Ultimately, persistent non-compliance by these and otherindividuals results in the prevalence of disease. This noncompliance isfrequently compounded by the emergence of drug-resistant strains ofmycobacteria. Effective compositions and vaccines that target variousstrains of mycobacteria are necessary to bring the increasing number oftuberculosis cases under control.

[0094] Chemotherapy is a standard treatment for tuberculosis. Somecurrent chemotherapy treatments require the use of three or four drugs,in combination, administered daily for two months, or administeredbiweekly for four to twelve months. TABLE 1 Treatment Schedules forStandard TB Drug Regimens. INDUC- CON- STAN- TION TINUATION DARD PHASEPHASE DRUG Dosing DURA- Dosing DURA- REGIMEN Schedule TION DRUG ScheduleTION Isoniazid Daily, 8 weeks Isoniazid 2/week, DOT 16 DOT weeksRifampicin Daily, 8 weeks Rifampicn 2/week, DOT 16 DOT weeks Pyrazin-Daily, 8 weeks amide DOT Ethambutol Daily, 8 weeks or DOT Streptomycin

[0095] Decades of misuse of existing antibiotics and poor compliancewith prolong and complex therapeutic regimens has led to mutations ofthe mycobacterium tuberculosis and has created an epidemic of drugresistance that threatens tuberculosis control world wide. The vastmajority of currently prescribed drugs, including the front line drugs,such as isoniazid, rifampin, pyrazinamide, ethambutol and streptomycinwere developed from the 1950s to the 1970s. Thus, this earlierdevelopment of tuberculosis chemotherapy did not have at its disposalthe implications of the genome sequence of Mycobacterium tuberculosis,the revolution in pharmaceutical drug discovery of the last decades, andthe use of national drug testing and combinational chemistry.

[0096] Consequently, the treatments of drug-resistant M. tuberculosisstrains, and latent tuberculosis infections, require newanti-tuberculosis drugs that provide highly effective treatments, andshortened and simplified tuberculosis chemotherapies. Moreover, it isdesirable that these drugs be prepared by a low-cost synthesis, sincethe demographics of the disease dictate that cost is a significantfactor.

[0097] The present invention provides methods and compositionscomprising a class of substituted ethylene diamine compounds effectivein treatment and prevention of disease caused by microorganismsincluding, but not limited to, bacteria. In particular, the methods andcompositions of the present invention are effective in inhibiting thegrowth of the microorganism, M. tuberculosis. The methods andcompositions of the present invention are intended for the treatment ofmycobacteria infections in human, as well as other animals. For example,the present invention may be particularly useful for the treatment ofcows infected by M. bovis.

[0098] As used herein, the term “tuberculosis” comprises disease statesusually associated with infections caused by mycobacteria speciescomprising M. tuberculosis complex. The term “tuberculosis” is alsoassociated with mycobacterial infections caused by mycobacteria otherthan M. tuberculosis (MOTT). Other mycobacterial species include M.avium-intracellulare, M. kansarii, M. fortuitum, M. chelonae, M. leprae,M. africanum, and M. microti, M. avium paratuberculosis, M.intracellulare, M. scrofulaceum, M. xenopi, M. marinum, M. ulcerans.

[0099] The present invention further comprises methods and compositionseffective for the treatment of infectious disease, including but notlimited to those caused by bacterial, mycological, parasitic, and viralagents. Examples of such infectious agents include the following:staphylococcus, streptococcaceae, neisseriaaceae, cocci,enterobacteriaceae, pseudomonadaceae, vibrionaceae, campylobacter,pasteurellaceae, bordetella, francisella, brucella, legionellaceae,bacteroidaceae, gram-negative bacilli, clostridium, corynebacterium,propionibacterium, gram-positive bacilli, anthrax, actinomyces,nocardia, mycobacterium, treponema, borrelia, leptospira, mycoplasma,ureaplasma, rickettsia, chlamydiae, systemic mycoses, opportunisticmycoses, protozoa, nematodes, trematodes, cestodes, adenoviruses,herpesviruses, poxyiruses, papovaviruses, hepatitis viruses,orthomyxoviruses, paramyxoviruses, coronaviruses, picornaviruses,reoviruses, togaviruses, flaviviruses, bunyaviridae, rhabdoviruses,human immunodeficiency virus and retroviruses.

[0100] The present invention further provides methods and compositionsuseful for the treatment of infectious disease, including by not limitedto, tuberculosis, leprosy, Crohn's Disease, aquired immunodeficiencysyndrome, lyme disease, cat-scratch disease, Rocky Mountain SpottedFever and influenza.

[0101] The anti-infective methods and compositions of the presentinvention contain one or more substituted ethylene diamine compounds. Inparticular, these compounds encompass a wide range of substitutedethylene diamine compounds having the following general formula:

[0102] where “R₁NH” is typically derived from a primary amine, and“R₂R₃N” is typically derived from a primary or secondary amine. Theethylene diamines of the present invention are prepared by a modularapproach using primary and secondary amines as building blocks, andcoupling the amine moieties with an ethylene linker building block.Representative primary amines, acyclic secondary amines, and cyclicsecondary amines are shown in FIGS. 2, 3, and 4, respectively.

[0103] Generally, chemical moieties R₁, R₂, and R₃ of the ethylenediamine compounds of the present invention are independently selectedfrom H, alkyl; aryl; alkenyl; alkynyl; aralkyl; aralkenyl; aralkynyl;cycloalkyl; cycloalkenyl; heteroalkyl; heteroaryl; halide; alkoxy;aryloxy; alkylthio; arylthio; silyl; siloxy; a disulfide group; a ureagroup; amino; and the like, including straight or branched chainderivatives thereof, cyclic derivatives thereof, substituted derivativesthereof, heteroatom derivatives thereof, heterocyclic derivativesthereof, functionalized derivatives thereof, salts thereof, such saltsincluding, but not limited to hydrochlorides and acetates, isomersthereof, or combinations thereof. For example, nitrogen-containingheterocyclic moieties include, but are not limited to, groups such aspyridinyl (derived from pyridine, and bonded through a ring carbon),piperidinyl (derived from piperidine and bonded through the ringnitrogen atom or a ring carbon), and pyrrolidinyl (derived frompyrrolidine and bonded through the ring nitrogen atom or a ring carbon).Examples of substituted, or functionalized, derivatives of R₁, R₂, andR₃ include, but are not limited to, moieties containing substituentssuch as acyl, formyl, hydroxy, acyl halide, amide, amino, azido, acid,alkoxy, aryloxy, halide, carbonyl, ether, ester, thioether, thioester,nitrile, alkylthio, arythio, sulfonic acid and salts thereof, thiol,alkenyl, alkynyl, nitro, imine, imide, alkyl, aryl, combinationsthereof, and the like. Moreover, in the case of alkylated derivatives ofthe recited moieties, the alkyl substituent may be pendant to therecited chemical moiety, or used for bonding to the amine nitrogenthrough the alkyl substituent.

[0104] Examples of chemical moieties R₁, R₂, and R₃ of the presentinvention include, but are not limited to: H; methyl; ethyl; propyl;butyl; pentyl; hexyl; heptyl; octyl; ethenyl; propenyl; butenyl;ethynyl; propynyl; butynyl; cyclopropyl; cyclobutyl; cyclopentyl;cyclohexyl; cyclooctyl cyclobutenyl; cyclopentenyl; cyclohexenyl;phenyl; tolyl; xylyl; benzyl; naphthyl; pyridinyl; furanyl;tetrahydro-1-napthyl; piperidinyl; indolyl; indolinyl; pyrrolidinyl;2-(methoxymethyl) pyrrolidinyl; piperazinyl; quinolinyl; quinolyl;alkylated-1,3-dioxolane; triazinyl; morpholinyl; phenyl pyrazolyl;indanyl; indonyl; pyrazolyl; thiadiazolyl; rhodaninyl; thiolactonyl;dibenzofuranyl; benzothiazolyl; homopiperidinyl; thiazolyl;quinonuclidinyl; isoxazolidinonyl; any isomers, derivatives, orsubstituted analogs thereof; or any substituted or unsubstitutedchemical species such as alcohol, ether, thiol, thioether, tertiaryamine, secondary amine, primary amine, ester, thioester, carboxylicacid, diol, diester, acrylic acid, acrylic ester, methionine ethylester, benzyl-1-cysteine ethyl ester, imine, aldehyde, ketone, amide, ordiene. Further examples of chemical moieties R₁, R₂, and R₃ of thepresent invention include, but are not limited to, the following speciesor substituted or alkylated derivatives of the following species,covalently bonded to the amine nitrogen: furan; tetrahydrofuran; indole;piperazine; pyrrolidine; pyrrolidinone; pyridine; quinoline; anthracene;tetrahydroquinoline; naphthalene; pyrazole; imidazole; thiophene;pyrrolidine; morpholine; and the like. One feature of the recitedspecies or substituted or alkylated derivatives of these species, isthat they may be covalently bonded to the amine nitrogen in any fashion,including through the pendant substituent or alkyl group, through theheteroatom as appropriate, or through a ring atom as appropriate, asunderstood by one of ordinary skill in the art.

[0105] The chemical moieties R₁, R₂, and R₃ of the present inventionalso include, but are not limited to, cyclic alkanes and cyclic alkenes,and include bridged and non-bridged rings. Examples of bridged ringsinclude, but are not limited to, the following groups: isopinocamphenyl;bornyl; norbornyl; adamantanetetyl; cis-(−)myrtanyl; adamantyl;noradamantyl; 6-azabicyclo[3.2.1]octane; exo-norbornane; and the like.

[0106] In one embodiment of the present invention, NR₂R₃ is derived froma cyclic secondary amine. Examples of a cyclic chemical moiety, NR₂R₃,of the present invention include, but are not limited to,4-benzyl-piperidine; 3-piperidinemethanol; piperidine; tryptamine;moropholine; 4-piperidinopiperidine; ethyl 1-piperazine carboxylate;1-(2-amino-ethyl)-piperazine; decahydroquinoline;1,2,3,4-tetrahydro-pyridoindole (reaction at either amine);3-amino-5-phenyl pyrazole; 3-aminopyrazole; 1-(2-fluorophenyl)piperazine; 1-proline methyl ester; histidinol; 1-piperonyl-piperazine;hexamethyleimine; 4-hydroxypiperidine; 2-piperidinemethanol; 1, 3,3-trimethyl-6-azabicyclo[3.2.1] octane; 3-pyrrolidinol;1-methylpiperazine; (S)-(+)-(2-pyrolidinylmethyl) pyrrolidine;1-methylhomopiperazine; 2-ethyl-piperidine; 1, 2, 3,4-tetrahydroisoquinoline; 1-(4-fluorophenyl) piperazine; d,1-tryptophanmethyl ester; tert-butyl (15,45)-(−)-2,5-diazabiclyclo[2.2.1]heptane-2-carboxylate; isonipecotamide;heptamethyleneimine; alpha-methyltryptamine; 6, 7-dimethoxy-1, 2, 3,4-tetrahydroisoquinoline; 3-aminopyrrolidine; 3, 5-dimethylpiperidine;2, 6-dimethylmorpholine; 1,4-dioxo-8-azaspiro[4.5]decane;1-methol-6,7-dihydroxy-1, 2, 3, 4-tetrahydroisoquinoline; 1, 3, 4, 6, 7,8-hexahydro-2H-pyrido (1,2-A) pyrimidine; 1, 2, 3,4-tetrahydroquinoline; 1-(2-methoxyphenyl) piperazine;1-(2-(2-hydroxyethoxy)ethyl) piperazine; (S)-(+)-2-(aminomethyl)pyrroli-dine; (3S(3a, 4Ab),8Ab)—N-t-butyl-D-ecahydro-3-isoquino-linecarboxamide; (R)-cycloserine;homopiperazine; 2, 6-dimethylpiperazine (reaction at either amine);iminodibenzyl; 5-methoxytryptamine; 4, 4′-bipiperidine;1-(2-hydroxyethyl) piperazine; 4-methylpiperidine; 1-histidine methylester; or methyl pipecoliate.

[0107] The R₁HN substituent is derived from a primary amine. The R₂R₃Nsubstituent is typically derived from a primary or secondary amine, butmay also arise from an amino acid, or an amino acid precursor. The aminoacid can transform into an amino alcohol. When an amino acid is employedas the source of the R₂R₃N moiety, the precursor compound may beselected from, among others, the following compounds and theirderivatives: d,1-tryptophan methyl ester; 1-methionine ethyl ester;1-lysine methyl ester (via reaction at either primary amine);(S)-benzyl-1-cysteine ethyl ester; 1-arginine methyl ester (via reactionat either primary amine); 1-glutamic acid ethyl ester; 1-histidinemethyl ester; or (3S (3a, 4Ab), 8A b)—N-t-butyl-D-ecahydro-3-iso-quinolinecarboxamide.

[0108] The R₄ moiety of the substituted ethylene diamine compounds ofthe present invention is typically selected from H, alkyl or aryl, butR₄ can also constitute alkenyl, alkynyl, aralkyl, aralkenyl, aralkynyl,cycloalkyl, cycloalkenyl, and the like. Examples of the R₄ chemicalmoiety include, but are not limited to: H; methyl; ethyl; propyl; butyl;pentyl; hexyl; heptyl; octyl; ethenyl; propenyl; butenyl; ethynyl;propynyl; butynyl; cyclobutyl; cyclopentyl; cyclohexyl; cyclobutenyl;cyclopentenyl; cyclohexenyl; phenyl; tolyl; xylyl; benzyl; naphthyl;straight or branched chain derivatives thereof; cyclic derivativesthereof; substituted, functionalized, and heteroatom derivativesthereof; and heterocyclic derivatives thereof, and the like. Typically,R₄ is selected from H, methyl, ethyl, butyl or phenyl. However, when R₄is “H” the ethylene diamine does not contain ethambutol.

[0109] A majority of the ethylene diamine compounds described hrein arepreferably prepared using a solid support synthesis, as set forth in oneof the representative reaction schemes shown in FIG. 1. However, when R₄is H, the reaction does not proceed well when sterically hindered aminesare used for R₁NH₂, or when diamines, such as amino alkylenemorpholine,or aminoalkylene-piperidines, are used for R₁NH₂. When R₄ is methyl, orphenyl, sterically hindered amines used for R₃R₂NH do not work well dueto steric hindrance at the reaction site. In this case, a competinghydrolysis reaction producing the corresponding amino alcohols, andincomplete reduction of the amidoethyleneamines, interfere with thereaction scheme. As a result, the desired diamine products form in lowyields.

[0110] The preparation of the ethylene diamines is preferablyaccomplished in six steps, using a rink-acid resin. The first step ofthe synthesis is converting the rink-acid resin to rink-chloride bytreatment with triphenylphosphine and hexachloroethane intetrahydrofuran (THF). This step is followed by addition of the primaryamine in the presence of Hunig's base (EtN(i-Pr)₂) in dichloroethane.The third step is the acylation of the resin-attached amine using eitherone of the two acylation routes shown in FIG. 1. The acylation step ispreferably accomplished using either α-chloroacetyl chloride,α-bromo-α-methyl acetylbromide, α-bromo-α-ethylacetyl bromide,α-bromo-α-butyl acetylbromide, or α-chloro-α-phenyl-acetylchloride, eachin the presence of pyridine in THF. Other acylation reagents known tothose skilled in the art may also be used, however, the α-bromoacetylhalides result in low product yields, which may be attributed to HBrelimination. The acylation may also be accomplished via a peptidecoupling mechanism using α-bromo-α-methylacetic acid, orα-chloro-α-methylacetic acid, in the presence ofbenzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate(PyBrop) and N₁N-diisopropylethyl amine (EtN(i-Pr)₂) in dichloromethane(DCM) and dimethylformamide (DMF). Again, other acylation reagents knownto those skilled in the art may also be used. The acylation step ispreferably performed twice to achieve better acylated product yields.

[0111] Introduction of the second nitrogen moiety is preferably achievedin the presence of Hunig's base in dimethylformamide (DMF). Reduction ofthe intermediate amine-amide is carried out using Red-Al (3.4M solutionof sodium bis (2-methoxyethoxy) aluminum hydride in toluene). The finalproduct is cleaved from the resin support using a 10% solution (byvolume) of trifluoroacetic acid (TFA) in dichloromethane (DCM). Thesolvent is evaporated, and the TFA salts of the final diamine productsare analyzed by mass spec, and screened against M. tuberculosis foreffectiveness. Some of the substituted ethylene diamines, prepared usingthe above-described solid-support synthesis, are also prepared using asolution phase synthesis described below.

[0112] Formation of the Substituted Ethylene Diamine Library

[0113] The solid support syntheses, shown in FIG. 1, are preferably usedto prepare a substituted ethylene diamine library. Solid phase synthesisoffers at least three principal advantages: (i) a reduced need forchromatographic procedures, (ii) the use of excess reagents to drive areaction forward in high yields, and (iii) the use of split and pooltechnologies for the synthesis of a large number of compounds. Solidsupport syntheses of 1, 2-diamine libraries have previously beenaccomplished by the reduction of short peptides (Cuervo et al., Peptides1994: Proceedings of the European Peptide Symposium; Maia HSL Ed., Esom:Leiden, 1995, 465-466). However, as described herein, an ethylenediamine library is created using amines, rather than simple amino acids,to allow for greater diversity in the building-block monomers. The firstthree steps of each support synthesis: the activation of the Rink-acidresin, the addition of the first amine, and the acylation step arecarried out in 10 ml tubes on a QUEST® 210 Synthesizer manufactured byARGONAUT TECHNOLOGIES®, Inc., Foster City, Calif. The synthesizerhandles up to twenty simultaneous reactions in 5 ml or 10 ml reactionvessels to allow for rapid synthesis of target compounds. Thesynthesizer provides programmable temperature control and agitation, andthe automated delivery of solvents into the reaction vessels. Theaddition of the second amine, the reduction with Red-A1, and thecleavage from the solid support are carried out in 2 ml wells in a96-well, chemically resistant plate.

[0114] Prior to the solid support synthesis, each amine, within numbers1 to 288, as shown in FIGS. 2, 3, and 4, is dissolved in DMF as a onemolar solution, and organized in three, 96-well plates (one amine perwell), to yield three master plates of these amines. An individualhaloacetyl amide from each primary amine and a particular R₄ group, isformed in the first three steps of the support synthesis. Individualhaloacetyl amides are then pooled into groups of ten or thirty. Asuspension of the pooled resins in a 2:1 mixture of DCM/THF is evenlydistributed into one, two or three reaction plates to assure 15-20 mg ofthe suspension per well. The number of reaction plates used is based onthe amount of suspension available. Each well of pooled resins isreacted with a corresponding amine from the master plates. FIG. 5provides a flow schematic for a representative pool. Each reactionoccurs in a separate well, in the presence of Hunig's base in DMF at70-75° C. for 16-20 hours. Each resulting amine-amide is reduced using65+w % Red-Al at room temperature. The reduction is followed by cleavagewith 10% vol. TFA in DCM. The solvents in each reaction well areevaporated, and the TFA salts of the diamines analyzed (mass spec), andscreened against M. tuberculosis. One plate of pooled diamines arescreened against M. smegmatis. Two randomly selected rows in each plate;i.e., 24 samples per 96-well plate, or 25% of the library, are examinedby mass spectroscopy. Specific protocols and detailed methods areprovided below in the Examples.

[0115] Screening Against M. tuberculosis

[0116] An entire library of synthesized substituted ethylene diamines(targeted number of compounds about 100,000), prepared as describedabove, was screened, in vitro, against M. tuberculosis in ethambutol(EMB) sensitive Luc-assay. The MIC (Minimum Inhibition Concentration)was also determined. The MIC is the minimum concentration of a growthinhibitor, here the substituted ethylene diamine, where there is nomultiplication of the microorganism under examination. Screening wasdone using a High-Throughout Screening (HTS) Luc assay with recombinantmycobacteria containing a promoter fusion of a luciferase to theEB-inducible gene (Luc assay). The Luc-assay and MIC assay are describedin detail below. These assays are well known to those skilled in theart. Based on this initial screening, 300+ compound mixtures showedanti-TB activity. FIG. 6 represents typical assay data in a luciferasereporter strain containing an Rv0341 EMB-inducible promoter. FIG. 6represents percent maximum Luminescense Count per Second (% Max. LCPS)for pooled compound mixtures in one row (row D) in one of the 96-wellplates.

[0117] Deconvolution of the Reactive Wells

[0118] The M. tuberculosis screening revealed approximately 300 activecompounds mixtures that were selected for deconvolution. In particular,wells possessing activity of approximately <12.5 μM in the HTS Lucassay, and/or an MIC of approximately <12.5 μM, were selected for atotal of 336 wells.

[0119] Deconvolutions were performed by discrete re-synthesis of eachsubstituted ethylene diamine compound in each active compound pool. Thepooled compounds in each active well were individually synthesized, andscreened. Syntheses of the targeted diamine compounds in each activepool were done in the 96-well plates using stored archived α-haloacetylamides (resin attached haloacetyl amides), according to the previouslydescribed reaction steps (the addition of the second amine, thereduction with Red-A1, and the cleavage from the solid support). Thearchived resins were stored as individual compounds at 4° C. The 96-wellplates were used for the remaining synthesis steps as previouslydescribed.

[0120] The same screening tests, MIC and HTS Luc assay, were performedon each deconvoluted compound. Representative Luminescence data fordeconvoluted compounds are shown in FIGS. 7 and 8. FIGS. 7 and 8represent the Luminescence Count per Second (LCPS) for individualcompounds.

[0121] Summary of Screening Results

[0122] Overall, the deconvolution screening results revealed about 2,000ethylene diamine compounds with inhibitory activity against M.tuberculosis. More than 150 of these compounds exhibited MICs equal toor lower than approximately 12.5 μM. FIG. 9 summarizes the MIC data forall synthesized discrete compounds with an MIC of 50 μM or less. FIG. 10summarizes Luc assay data for all compounds that exhibit at least 10%activity at each concentration (the results are not cumulative). The MICand Luc activities were obtained for non-purified samples, with chemicalyields of approximately 20%, based on an assumed 80% yield at eachreaction step. In the Luc assay, 32 compounds exhibited activity at 1.56μM, and in the MIC assay, at least 11 compounds had an MIC of 3.13 μM.

[0123] The total frequency of the top thirteen amines that contributedto the activity of the substituted ethylene diamines are shown in FIG.11, with each amine represented by its numerical designation. Theseamines include the following:

[0124] #11 2,3-Dimethylcylochexy amines

[0125] #18 3,3-Diphenylpropylamine

[0126] #44 1-Adamantanemethylamine

[0127] #47 2,2-Diphenylethylamine

[0128] #63 (S)-2-Amino-1-butanol

[0129] #74.1 (−)-cis-Myrtanylamine

[0130] #77.1 Cyclooctylamine

[0131] #78.1 2-Adamantamine

[0132] #105a (1R,2R,3R,5 S)-(−)-Isopinocampheylamine

[0133] #231 2-Methoxyphenethylamine

[0134] #255 (S)-Cylcohexylethylamine

[0135] #266 Undecylamine

[0136] #272 Geranylamine

[0137] Other amines that contributed to the activity of the substitutedethylene diamines are shown in Table 2. The compounds in Table 2 aresorted by their MIC results. Some compounds, synthesized in largerquantities (2-60 mg) on the Quest® Synthesizer, and purified by HPLCusing semi-preparative C18-column, are shown in Table 3. Generally, thefinal purity of each compound in Table 3 was at least 90%. TABLE 2Synthetic Substituted Diethylene Diamines Sorted by Minimum InhibitionConcentration MIC % N1 N2 R4 (μM) Induction 3,3-Diphenylpropylamineexo-Aminonorbornane Hydrogen 3.13 53.70 2,2-Diphenylamine(+)-Isopinocampheylamine Hydrogen 3.13 93.94 2,2-Diphenylaminecis-(−)-Myrtanylamine Hydrogen 3.13 64.49 2,2-DiphenylamineCyclooctylamine Hydrogen 3.13 63.44 2,2-Diphenylamine3,4-Dihydroxynorephedrine Hydrogen 3.13 42.80 5-AminoquinolineCyclohexylamine Hydrogen 3.13 18.33 5-Aminoquinoline tert-OctylamineHydrogen 3.13 20.85 5-Aminoquinoline 4-Methylcyclohexylamine Hydrogen3.13 26.33 cis-(−)-Myrtanylamine (+)-Bornylamine Hydrogen 3.13 100.00cis-(−)-Myrtanylamine 1-Adamantanemethylamine Hydrogen 3.13 85.20cis-(−)-Myrtanylamine (−)-Isopinocampheylamine Hydrogen 3.13 60.941-Adamantanemethylamine tert-Octylamine Hydrogen 4.7 9.813,4-Dimethoxyphenethylamine 1-Adamantanemethylamine Hydrogen 6.25 11.453,4-Dimethoxyphenethylamine Hexetidine (mixture of isomers) Hydrogen6.25 0 3,4-Dimethoxyphenethylamine Dehydroabietylamine Hydrogen 6.25 03,3-Diphenylpropylamine 1-Adamantanemethylamine Hydrogen 6.25 9.533,3-Diphenylpropylamine 2-Methylcyclohexylamine Hydrogen 6.25 50.08 (mixof cis and trans) 3,3-Diphenylpropylamine 1,3-DimethylbutylamineHydrogen 6.25 39.40 3,3-Diphenylpropylamine 1-(1-Adamantyl)ethylamine,Hydrogen 6.25 45.14 HCl 3,3-Diphenylpropylamine(S)-(−)-Cyclohexylethylamine Hydrogen 6.25 43.49 3,3-Diphenylpropylamine(R)-(−)-Cyclohexylethylamine Hydrogen 6.25 34.54 3,3-Diphenylpropylamine1-Adamantanemethylamine Methyl 6.25 16.14 Propylamine Hexetidine(mixture of isomers) Hydrogen 6.25 0 Phenethylamine Hexetidine (mixtureof isomers) Hydrogen 6.25 0 b-Methylphenethylamine Hexetidine (mixtureof isomers) Hydrogen 6.25 0 b-Methylphenethylamine Undecylamine Hydrogen6.25 0 2,2-Diphenylamine (+)-Bornylamine Hydrogen 6.25 87.862,2-Diphenylamine (−)-Isopinocampheylamine Hydrogen 6.25 77.802,2-Diphenylamine alpha-Methyltryptamine Hydrogen 6.25 55.072,2-Diphenylamine alpha-Methyltryptamine Hydrogen 6.25 23.082,2-Diphenylamine 4-Phenylbutylamine Hydrogen 6.25 2,2-Diphenylamine2,5-Dimethoxyphenethylamine Hydrogen 6.25 2,2-Diphenylamine2,4-Dichlorophenethylamine Hydrogen 6.25 2,2-Diphenylamine2-(2-Aminomethyl) Hydrogen 6.25 phenylthio)benzyl alcohol2,2-Diphenylamine 1-(1-Naphthyl)ethylamine Hydrogen 6.25 7.20 Veratrylamine 2,5-Dimethoxyphenethylamine Hydrogen 6.25 Veratryl amine2-(2-Aminomethyl) Hydrogen 6.25 phenylthio)benzyl alcohol5-Aminoquinoline 2-Aminoheptane Hydrogen 6.25 26.22 5-Aminoquinoline1-Adamantanamine Hydrogen 6.25 18.91 1-Aminomethyl-1- Hexetidine(mixture of isomers) Hydrogen 6.25 cyclohexanol, HClcis-(−)-Myrtanylamine 2,3-Dimethylcyclohexylamine Hydrogen 6.25 100.00cis-(−)-Myrtanylamine 3,3-Diphenylpropylamine Hydrogen 6.25 87.78cis-(−)-Myrtanylamine (+)-Isopinocampheylamine Hydrogen 6.25 93.10cis-(−)-Myrtanylamine 2,2-Diphenylamine Hydrogen 6.25 81.84cis-(−)-Myrtanylamine cis-(−)-Myrtanylamine Hydrogen 6.25 68.24cis-(−)-Myrtanylamine 1,3,3-Trimethyl-6- Hydrogen 6.25 68.18azabicyclo[3.2.1]octane cis-(−)-Myrtanylamine 1-AdamantanemethylamineMethyl 6.25 24.22 cis-(−)-Myrtanylamine cis-(−)-Myrtanylamine Methyl6.25 44.14 Cyclooctylamine 3,3-Diphenylpropylamine Hydrogen 6.25 100.00Cyclooctylamine (−)-Isopinocampheylamine Hydrogen 6.25 59.13sec-Butylamine Hexetidine (mixture of isomers) Hydrogen 6.253-Methylbenzylamine Hexetidine (mixture of isomers) Hydrogen 6.253-Methylbenzylamine Undecylamine Hydrogen 6.25 2-MethoxyethylamineHexetidine (mixture of isomers) Hydrogen 6.25 Geranylamine2-Adamantanamine, HCl Hydrogen 6.25 25.66 1-Adamantanemethylamine4-Benzylpiperidine Hydrogen 9.4 0 i-Adamantanemethylamine2,3-Dimethylcyclohexylamine Hydrogen 9.4 0 1-Adamantanemethylamine3,3-Diphenylpropylamine Hydrogen 9.4 40.06 1-Adamantanemethylamine1-Adamantanemethylamine Hydrogen 9.4 15.25 1-Adamantanemethylamine2,2-Diphenylamine Hydrogen 9.4 0 1-Adamantanemethylamine1,3,3-Trimethyl-6- azabicyclo[3.2.1]octane Hydrogen 9.4 01-Adamantanemethylamine 138 Hydrogen 9.4 0 3-Phenyl-1-propylamine 138Hydrogen 9.4 2,2-Diphenylamine 1-Adamantanemethylamine Hydrogen 9.465.89 2,2-Diphenylamine 138 Hydrogen 9.4 Furfurylamine Hexetidine(mixture of isomers) Hydrogen 12.5 0 3,4,5-TrimethoxybenzylamineHexetidine (mixture of isomers) Hydrogen 12.5 01-Methyl-3-phenylpropylamine Dehydroabietylamine Hydrogen 12.5 0Cyclobutylamine Hexetidine (mixture of isomers) Hydrogen 12.5 02-Fluorobenzylamine Hexetidine (mixture of isomers) Hydrogen 12.5 02-Fluorobenzylamine Dehydroabietylamine Hydrogen 12.5 03,4-Dimethoxyphenethylamine Undecylamine Hydrogen 12.5 03,3-Diphenylpropylamine exo-Aminonorbornane Hydrogen 12.5 14.383,3-Diphenylpropylamine Decahydroquinoline Hydrogen 12.5 22.523,3-Diphenylpropylamine Hexetidine (mixture of isomers) Hydrogen 12.5 03,3-Diphenylpropyl amine 4-Phenylbutylamine Hydrogen 12.5 03,3-Diphenylpropylamine 2-Methoxyphenethylamine Hydrogen 12.5 6.823,3-Diphenylpropylamine 2,4-Dichlorophenethylamine Hydrogen 12.5 03,3-Diphenylpropylamine 1-Aminoindan Hydrogen 12.5 18.053,3-Diphenylpropylamine Undecylamine Hydrogen 12.5 03,3-Diphenylpropylamine Dehydroabietylamine Hydrogen 12.5 03,3-Diphenylpropylamine 2-(1-Cyclohexenyl)ethylamine Methyl 12.5 9.53,3-Diphenylpropylamine cis-(−)-Myrtanylamine Methyl 12.5 18.413,3-Diphenylpropylamine Cyclooctylamine Methyl 12.5 20.84 PropylamineDehydroabietylamine Hydrogen 12.5 0 Phenethylamine DehydroabietylamineHydrogen 12.5 0 Cyclohexylamine Hexetidine (mixture of isomers) Hydrogen12.5 0 3-Amino-1-propanol Hexetidine (mixture of isomers) Hydrogen 12.50 b-Methylphenethylamine Dehydroabietylamine Hydrogen 12.5 04-Methoxyphenethylamine 2-Fluorophenethylamine Hydrogen 12.5 04-Methoxyphenethylamine 2-(1-Cyclohexenyl)ethylamine Hydrogen 12.5 04-Methoxyphenethylamine 2,4-Dimethoxybenzylamine Hydrogen 12.5 04-Methoxphenethylamine 4-Fluorophenethylamine Hydrogen 12.5 16.784-Methoxyphenethylamine Hexetidine (mixture of isomers) Hydrogen 12.5 0Tetrahydrofurfurylamine Hexetidine (mixture of isomers) Hydrogen 12.5 0Amylamine 4-Fluorophenethyl amine Hydrogen 12.5 0 3-Phenyl-1-propylamine2-(1-Cyclohexenyl)ethylamine Hydrogen 12.5 3-Phenyl-1-propylamine4-Fluorophenethylamine Hydrogen 12.5 12.94 2,2-Diphenylaminetert-Amylamine Hydrogen 12.5 9.05 2,2-Diphenylamine UndecylamineHydrogen 12.5 2,2-Diphenylamine Dehydroabiethylamine Hydrogen 12.52,2-Diphenylamine cis-(−)-Myrtanylamine Methyl 12.5 45.181-(3-Aminopropyl)-2- 2,5-Dimethoxyphenethylamine Hydrogen 12.5pyrrolidinone (tech) 1-(3-Aminopropyl)-2-2-(2-Aminomethyl)phenylthio)benzyl Hydrogen 12.5 pyrrolidinone (tech)alcohol 4-(Trifluoromethyl)benzylamine 2,5-DimethoxyphenethylamineHydrogen 12.5 4-(Trifluoromethyl)benzylamine 1-(1-Naphthyl)ethylamineHydrogen 12.5 Veratryl amine 4-Phenylbutylamine Hydrogen 12.55-Amino-1-pentanol 2,5-Dimethoxyphenethylamine Hydrogen 12.55-Amino-1-pentanol 2-(2-Aminomethyl)phenylthio)benzyl Hydrogen 12.5alcohol 2-(1-Cyclohexenyl)ethylamine 2-(1-Cyclohexenyl)ethylamineHydrogen 12.5 2-(1-Cyclohexenyl)ethylamine 4-FluorophenethylamineHydrogen 12.5 2-(1-Cyclohexenyl)ethylamine 4-Phenylbutylamine Hydrogen12.5 2-(1-Cyclohexenyl)ethylamine 2,5-Dimethoxyphenethylamine Hydrogen12.5 2-(2-Aminomethyl) 2-(1-Cyclohexenyl)ethylamine phenylthio)benzylalcohol Hydrogen 12.5 1-Aminomethyl-1- 2,5-DimethoxyphenethylamineHydrogen 12.5 cyclohexanol, HCl 3-Fluorobenzylamine2,5-Dimethoxyphenethylamine Hydrogen 12.5 4-Amino-1-butanol Hexetidine(mixture of isomers) Hydrogen 12.5 2-Ethoxybenzylamine Hexetidine(mixture of isomers) Hydrogen 12.5 cis-(−)-Myrtanylamine CyclooctylamineHydrogen 12.5 67.73 cis-(−)-Myrtanylamine 4-MethylcyclohexylamineHydrogen 12.5 18.39 cis-(−)-Myrtanylamine 1-Adamantanamine Hydrogen 12.560.16 cis-(−)-Myrtanylamine 3,3-Diphenylpropylamine Methyl 12.5 22.32Cyclooctylamine (+)-Isopinocampheylamine Hydrogen 12.5 57.83Cyclooctylamine (+)-Bornylamine Hydrogen 12.5 100.00 Cyclooctylamine1-Adamantanemethylamine Hydrogen 12.5 52.95 Cyclooctylamine2,2-Diphenylamine Hydrogen 12.5 71.43 Cyclooctylaminecis-(−)-Myrtanylamine Hydrogen 12.5 84.56 CyclooctylamineCyclooctylamine Hydrogen 12.5 59.21 Cyclooctylamine Hexetidine (mixtureof isomers) Hydrogen 12.5 Cyclooctylamine Aminodiphenylmethane Hydrogen12.5 Cyclooctylamine Undecylamine Hydrogen 12.5 5.61 Cyclooctylamine3,3-Diphenylpropylamine Methyl 12.5 53.92 Cyclooctylamine(+)-Isopinocampheylamine Methyl 12.5 Cyclooctylaminecis-(−)-Myrtanylamine Methyl 12.5 33.89 4-ChlorophenylalaninolHexetidine (mixture of isomers) Hydrogen 12.5 (−)-Isopinocampheylamine3,3-Diphenylpropylamine Hydrogen 12.5 23.68 (−)-Isopinocampheylamine(+)-Bornylamine Hydrogen 12.5 44.85 (−)-Isopinocampheylamine2-Amino-1-propanol, d,l Hydrogen 12.5 46.19 (−)-Isopinocampheylaminecis-(−)-Myrtanylamine Hydrogen 12.5 33.87 (−)-Isopinocampheylamine2-Adamantanamine, HCl Hydrogen 12.5 24.29 (−)-IsopinocampheylamineAminodiphenylmethane Hydrogen 12.5 48.35 Allamine Hexetidine (mixture ofisomers) Hydrogen 12.5 3-Ethoxypropylamine Hexetidine (mixture ofisomers) Hydrogen 12.5 sec-Butylamine Dehydroabietylamine Hydrogen 12.52-Aminoheptane Dehydroabietylamine Hydrogen 12.5 Ethanolamine Hexetidine(mixture of isomers) Hydrogen 12.5 3-Methylbenzylamine4-Phenylbutylamine Hydrogen 12.5 3-Methylbenzylamine2,4-Dichlorophenethylamine Hydrogen 12.5 3-MethylbenzylamineDehydroabietylamine Hydrogen 12.5 Piperonylamine Hexetidine (mixture ofisomers) Hydrogen 12.5 Piperonylamine Dehydroabietylamine Hydrogen 12.52-Methoxyethyl amine Dehydroabietyl amine Hydrogen 12.54-Fluorophenethylamine Hexetidine (mixture of isomers) Hydrogen 12.53-o-Methyldopamine, HCl Hexetidine (mixture of isomers) Hydrogen 12.53-o-Methyldopamine, HCl Undecylamine Hydrogen 12.5 3-o-Methyldopamine,HCl Dehydroabietylamine Hydrogen 12.5 3-Fluorophenethylamine Hexetidine(mixture of isomers) Hydrogen 12.5 3-FluorophenethylamineDehydroabietylamine Hydrogen 12.5 2-Methoxyphenethylamine Hexetidine(mixture of isomers) Hydrogen 12.5 2-MethoxyphenethylamineAminodiphenylmethane Hydrogen 12.5 34.67 2-Fluoroethylamine, HClHexetidine (mixture of isomers) Hydrogen 12.5 2-Amino-1-phenylethanolHexetidine (mixture of isomers) Hydrogen 12.5 2-Amino-1-phenylethanolDehydroabietylamine Hydrogen 12.5 2,5-Dimethoxyphenethylamine2-Adamantanamine, HCl Hydrogen 12.5 22.18 2-(2-Chlorophenyl)ethylamineN-Allylcyclopentylamine Hydrogen 12.5 62.31 2-(2-Chlorophenyl)ethylamineHexetidine (mixture of isomers) Hydrogen 12.5 3-HydroxytyramineHexetidine (mixture of isomers) Hydrogen 12.5 4- 2-Adamantanamine, HClHydrogen 12.5 28.34 (Trifluoromethoxy)benzylamine Geranylamine(+)-Bornylamine Hydrogen 12.5 Geranylamine 1,3,3-Trimethyl-6- Hydrogen12.5 37.42 azabicyclo[3.2.1]octane Geranylamine 2-EthylpiperidineHydrogen 12.5 29.81 Geranylamine 1-Adamantanamine Hydrogen 12.5 16.63Geranylamine N-Allylcyclopentylamine Hydrogen 12.5 74.86 GeranylamineAminodiphenylmethane Hydrogen 12.5 57.93 GeranylamineDehydroabietylamine Hydrogen 12.5 1-AdamantanemethylamineDecahydroquinoline Hydrogen 18.8 0 1-Adamantanemethylamine1-Adamantanamine Hydrogen 18.8 0 2,2-Diphenylamine2,3-Dimethylcyclohexylamine Hydrogen 18.8 23.60 2,2-Diphenylaminetert-Octylamine Hydrogen 18.8 19.29 2,2-Diphenylamine DecahydroquinolineHydrogen 18.8 8.96 4-Methylbenzylamine Furfurylamine Hydrogen 25 13.464-Methylbenzylamine Benzylamine Hydrogen 25 17.07 4-MethylbenzylamineHexetidine (mixture of isomers) Hydrogen 25 0 4-MethylbenzylamineDehydroabietylamine Hydrogen 25 0 Cyclopentylamine Hexetidine (mixtureof isomers) Hydrogen 25 0 Cyclopentylamine Dehydroabietylamine Hydrogen25 0 Furfurylamine Furfurylamine Hydrogen 25 01-Methyl-3-phenylpropylamine Hexetidine (mixture of isomers) Hydrogen 250 1-Methyl-3-phenylpropylamine Undecylamine Hydrogen 25 01,2,3,4-Tetrahydro-1- Undecylamine Hydrogen 25 6.24 naphthylamine1,2,3,4-Tetrahydro-1- Dehydroabietylamine Hydrogen 25 0 naphthylamine2,3-Dimethylcyclohexylamine Undecylamine Hydrogen 25 02,3-Dimethylcyclohexylamine Dehydroabietylamine Hydrogen 25 0 TyramineHexetidine (mixture of isomers) Hydrogen 25 0 Tyramine UndecylamineHydrogen 25 0 Tyramine Dehydroabietylamine Hydrogen 25 0 Tyraminecis-(−)-Myrtanylamine Methyl 25 0 2-Fluorobenzylamine UndecylamineHydrogen 25 0 (R)-2-Amino-1-butanol Hexetidine (mixture of isomers)Hydrogen 25 0 3,3-Diphenylpropylamine (S)-(+)-1-Amino-2-propanolHydrogen 25 0 3,3-Diphenylpropylamine 2-Ethylpiperidine Hydrogen 2511.32 3,3-Diphenylpropylamine N-Allylcyclopentylamine Hydrogen 25 11.633,3-Diphenylpropylamine Aminodiphenylmethane Hydrogen 25 03,3-Diphenylpropylamine 3,5-Dimethylpiperidine (cis- Hydrogen 25 30.28and trans-) 3,3-Diphenylpropylamine Allylcyclohexylamine Hydrogen 259.10 Propylamine Undecylamine Hydrogen 25 0 Phenethylamine UndecylamineHydrogen 25 0 Tryptamine (S)-(+)-1-Amino-2-propanol Hydrogen 25 0Tryptamine 2-Amino-2-methyl-1-propanol Hydrogen 25 0 CyclohexylamineUndecylamine Hydrogen 25 0 Cyclohexylamine Dehydroabietylamine Hydrogen25 0 (+)-Isopinocampheylamine Dehydroabietylamine Hydrogen 25 0Benzylamine Hexetidine (mixture of isomers) Hydrogen 25 BenzylamineUndecylamine Hydrogen 25 3-Amino-1-propanol Dehydroabietylamine Hydrogen25 0 2-Fluorophenethylamine 2-Fluorophenethylamine Hydrogen 25 02-Fluorophenethylamine Veratryl amine Hydrogen 25 02-Fluorophenethylamine 2,4-Dimethoxybenzylamine Hydrogen 25 02-Fluorophenethylamine 2-Amino-2-methyl-1-propanol Hydrogen 25 02-Fluorophenethylamine 4-Fluorophenethylamine Hydrogen 25 02-Fluorophenethylamine Hexetidine (mixture of isomers) Hydrogen 25 02-Fluorophenethylamine 1-(1-Naphthyl)ethylamine Hydrogen 25 02-Fluorophenethylamine 1-Adamantanemethylamine Methyl 25 3.212-Fluorophenethylamine cis-(−)-Myrtanylamine Methyl 25 4.89b-Methylphenethylamine 4-Phenylbutylamine Hydrogen 25 0b-Methylphenethylamine 2,4-Dichlorophenethylamine Hydrogen 25 0b-Methylphenethylamine 1-(1-Naphthyl)ethylamine Hydrogen 25 04-Methoxyphenethylamine 1-Adamantanemethylamine Hydrogen 25 04-Methoxyphenethylamine 1-(3-Aminopropyl)-2- Hydrogen 25 0 pyrrolidinone(tech) 4-Methoxyphenethylamine Veratryl amine Hydrogen 25 04-Methoxyphenethylamine Undecylamine Hydrogen 25 04-Methoxyphenethylamine Dehydroabietylamine Hydrogen 25 0Tetrahydrofurfurylamine Dehydroabietylamine Hydrogen 25 0 Amylamine2-Fluorophenethylamine Hydrogen 25 0 Amylamine2-(1-Cyclohexenyl)ethylamine Hydrogen 25 0 Amylamine2,4-Dimethoxybenzylamine Hydrogen 25 0 3-Phenyl-1-propylamine2-Fluorophenethylamine Hydrogen 25 3-Phenyl-1-propylamine1-Adamantanemethylamine Hydrogen 25 3-Phenyl-1-propylamine2,4-Dimethoxybenzylamine Hydrogen 25 3-Phenyl-1-propylamine Hexetidine(mixture of isomers) Hydrogen 25 3-Phenyl-1-propylamine4-Phenylbutylamine Hydrogen 25 3-Phenyl-1-propylamine2,4-Dichlorophenethylamine Hydrogen 25 3-Phenyl-1-propylamineUndecylamine Hydrogen 25 3-Phenyl-1-propylamine DehydroabietylamineHydrogen 25 2,2-Diphenylamine 4-(2-Aminoethyl)morpholine Hydrogen 252,2-Diphenylamine 1-(3-Aminopropyl)-2- Hydrogen 25 pyrrolidinone (tech)2,2-Diphenylamine 2-(1-Cyclohexenyl)ethylamine Hydrogen 252,2-Diphenylamine 2,4-Dimethoxybenzylamine Hydrogen 25 2,2-Diphenylamine4-(3-Aminopropyl)morpholine Hydrogen 25 2,2-Diphenylamine4-Fluorophenethylamine Hydrogen 25 2,2-Diphenylamine Hexetidine (mixtureof isomers) Hydrogen 25 2,2-Diphenylamine (S)-(−)-CyclohexylethylamineHydrogen 25 2,2-Diphenylamine 1-Adamantanemethylamine Methyl 25 5.841-(3-Aminopropyl)-2- 4-Phenylbutylamine Hydrogen 25 pyrrolidinone (tech)4-(Trifluoromethyl)benzylamine 1-Adamantanemethylamine Hydrogen 254-(Trifluoromethyl)benzylamine tert-Amylamine Hydrogen 254-(Trifluoromethyl)benzylamine alpha-Methyltryptamine Hydrogen 25 6.064-(Trifluoromethyl)benzylamine 4-Phenylbutylamine Hydrogen 254-(Trifluoromethyl)benzylamine 2-(2-Aminomethyl) Hydrogen 25 5.13phenylthio)benzyl alcohol 4-(Trifluoromethyl)benzylamine UndecylamineHydrogen 25 4-(Trifluoromethyl)benzylamine (−)-3,4-DihydroxynorephedrineHydrogen 25 4-(Trifluoromethyl)benzylamine Dehydroabietylamine Hydrogen25 Veratryl amine tert-Amylamine Hydrogen 25 5-Amino-1-pentanol4-Phenylbutylamine Hydrogen 25 2-(1-Cyclohexenyl)ethylamine2-Fluorophenethylamine Hydrogen 25 2-(1-Cyclohexenyl)ethylamine1-Adamantanemethylamine Hydrogen 25 1-Aminomethyl-1- 4-PhenylbutylamineHydrogen 25 cyclohexanol, HCl 3-Fluorobenzylamine 4-PhenylbutylamineHydrogen 25 3-Fluorobenzylamine 2-(2-Aminomethyl)phenylthio)benzylHydrogen 25 alcohol 2,4-Dimethoxybenzylamine 1-Adamantanamine Hydrogen25 2,4-Dimethoxybenzylamine Hexetidine (mixture of isomers) Hydrogen 252,4-Dimethoxybenzylamine Undecylamine Hydrogen 252,4-Dimethoxybenzylamine Dehydroabietylamine Hydrogen 252-Ethoxybenzylamine 1-Adamantanamine Hydrogen 25 2-EthoxybenzylamineN-Phenylethyldiamine Hydrogen 25 2-Ethoxybenzylamine2,4-Dichlorophenethylamine Hydrogen 25 2-Ethoxybenzylamine2-(2-Chlorophenyl)ethylamine Hydrogen 25 3.89 2-EthoxybenzylamineUndecylamine Hydrogen 25 2-Ethoxybenzylamine DehydroabietylamineHydrogen 25 cis-(−)-Myrtanylamine 2-(1-Cyclohexenyl)ethylamine Hydrogen25 cis-(−)-Myrtanylamine Hexetidine (mixture of isomers) Hydrogen 25cis-(−)-Myrtanylamine Aminodiphenylmethane Hydrogen 25cis-(−)-Myrtanylamine 2,4-Dichlorophenethylamine Hydrogen 25cis-(−)-Myrtanylamine (S)-(−)-Cyclohexylethylamine Hydrogen 25 28.94cis-(−)-Myrtanylamine Undecylamine Hydrogen 25 cis-(−)-Myrtanylamine(+)-Isopinocampheylamine Methyl 25 cis-(−)-Myrtanylamine CyclooctylamineMethyl 25 24.92 Cyclooctylamine 2,3-Dimethylcyclohexylamine Hydrogen 2550.55 Cyclooctylamine (S)-2-Amino-1-butanol Hydrogen 25 100.00Cyclooctylamine 2-Adamantanamine, HCl Hydrogen 25 29.61 Cyclooctylamine4-Phenylbutylamine Hydrogen 25 Cyclooctylamine 2-ChlorobenzylamineHydrogen 25 Cyclooctylamine 2-Aminoindan, HCl Hydrogen 25Cyclooctylamine Dehydroabietylamine Hydrogen 25 Cyclooctylamine1-(1-Naphthyl)ethylamine Hydrogen 25 4.62 Cyclooctylamine1-Adamantanemethylamine Methyl 25 14.20 2,3-DimethoxybenzylamineHexetidine (mixture of isomers) Hydrogen 25 2,3-DimethoxybenzylamineUndecylamine Hydrogen 25 2,3-Dimethoxybenzylamine DehydroabietylamineHydrogen 25 4-Methylcyclohexylamine Hexetidine (mixture of isomers)Hydrogen 25 4-Methylcyclohexylamine Undecylamine Hydrogen 254-Methylcyclohexylamine Dehydroabietylamine Hydrogen 254-Fluorobenzylamine Dibenzylamine Hydrogen 25 27.98 trans-2-Cyclooctylamine Hydrogen 25 32.80 Phenylcyclopropylamine, HCl trans-2-2-Adamantanamine, HCl Hydrogen 25 18.99 Phenylcyclopropylamine, HCltrans-2- 1-Adamantanamine Hydrogen 25 18.84 Phenylcyclopropylamine, HClThiomicamine Hexetidine (mixture of isomers) Hydrogen 25(R)-1-Amino-2-propanol Hexetidine (mixture of isomers) Hydrogen 254-Chlorophenylalaninol 2,4-Dichlorophenethylamine Hydrogen 254-Chlorophenylalaninol Undecylamine Hydrogen 25 4-ChlorophenylalaninolDehydroabietylamine Hydrogen 25 I-Leucinol Hexetidine (mixture ofisomers) Hydrogen 25 I-Leucinol 2,4-Dichlorophenethylamine Hydrogen 25I-Leucinol Dehydroabietylamine Hydrogen 25 (−)-Isopinocampheylamine2-Methoxyphenethylamine Hydrogen 25 29.59 (−)-IsopinocampheylamineUndecylamine Hydrogen 25 Allylamine Dehydroabietylamine Hydrogen 253-Amino-1,2-propanediol Hexetidine (mixture of isomers) Hydrogen 253-Ethoxypropylamine 3,3-Diphenylpropylamine Hydrogen 253-Ethoxypropylamine Undecylamine Hydrogen 25 3-EthoxypropylamineDehydroabietylamine Hydrogen 25 sec-Butylamine2,4-Dichlorophenethylamine Hydrogen 25 sec-Butylamine UndecylamineHydrogen 25 2-Aminoheptane Hexetidine (mixture of isomers) Hydrogen 252-Aminoheptane 4-Phenylbutylamine Hydrogen 25 2-Aminoheptane2,4-Dichlorophenethylamine Hydrogen 25 1-NaphthalenemethylamineHexetidine (mixture of isomers) Hydrogen 25 1-Naphthalenemethylamine4-Phenylbutylamine Hydrogen 25 1-Naphthalenemethylamine2,4-Dichlorophenethylamine Hydrogen 25 1-NaphthalenemethylamineUndecylamine Hydrogen 25 Ethanolamine Dehydroabietylamine Hydrogen 25Piperonylamine 4-Phenylbutylamine Hydrogen 25 1-EthylpropylamineHexetidine (mixture of isomers) Hydrogen 25 1-EthylpropylamineDehydroabietylamine Hydrogen 25 Isopropylamine Hexetidine (mixture ofisomers) Hydrogen 25 4-Fluorophenethylamine 4-Phenylbutylamine Hydrogen25 4-Fluorophenethylamine 2,4-Dichlorophenethylamine Hydrogen 254-Fluorophenethylamine Dehydroabietylamine Hydrogen 253-Fluorophenethylamine Undecylamine Hydrogen 25 2-Thiopheneethylamine2-Adamantanamine, HCl Hydrogen 25 19.09 2-Methylcyclohexylamine (mixHexetidine (mixture of isomers) Hydrogen 25 of cis and trans)2-Methylcyclohexylamine (mix Dehydroabietylamine Hydrogen 25 of cis andtrans) 2-Methoxyphenethylamine 2-Adamantanamine, HCl Hydrogen 25 26.772-Methoxyphenethylamine (−)-Isopinocampheylamine Hydrogen 25 31.952-Methoxyphenethylamine 1-Adamantanamine Hydrogen 25 24.382-Methoxyphenethylamine N-Allylcyclopentylamine Hydrogen 25 14.562-Methoxyphenethylamine 4-Phenylbutylamine Hydrogen 252-Methoxyphenethylamine Undecylamine Hydrogen 25 2-MethoxyphenethylamineDehydroabietylamine Hydrogen 25 2-Fluoroethylamine, HCl UndecylamineHydrogen 25 2-Fluoroethylamine, HCl Dehydroabietylamine Hydrogen 252-Aminoindan, HCl 2-Adamantanamine, HCl Hydrogen 25 17.722-Amino-1-phenylethanol Undecylamine Hydrogen 252,5-Dimethoxyphenethylamine (+)-Bornylamine Hydrogen 25 25.782,5-Dimethoxyphenethylamine Noradamantamine, HCl Hydrogen 25 11.732,5-Dimethoxyphenethylamine 1-Adamantanamine Hydrogen 25 12.572-(2-Chlorophenyl)ethylamine 4-Phenylbutylamine Hydrogen 252-(2-Chlorophenyl)ethylamine Undecylamine Hydrogen 252-(2-Chlorophenyl)ethylamine 1-(1-Naphthyl)ethylamine Hydrogen 252-(2-Aminomethyl)phenylthio)benzyl Hexetidine (mixture of isomers)Hydrogen 25 alcohol 2-(2-Aminomethyl)phenylthio)benzyl4-Phenylbutylamine Hydrogen 25 alcohol2-(2-Aminomethyl)phenylthio)benzyl Undecylamine Hydrogen 25 alcohol1-Aminoindan Hexetidine (mixture of isomers) Hydrogen 25 1-AminoindanUndecylamine Hydrogen 25 1-Aminoindan Dehydroabietylamine Hydrogen 251,3-Dimethylbutylamine Hexetidine (mixture of isomers) Hydrogen 251,3-Dimethylbutylamine Undecylamine Hydrogen 25 5.921,3-Dimethylbutylamine Dehydroabietylamine Hydrogen 25(S)-(−)-Cyclohexylethylamine (−)-Isopinocampheylamine Hydrogen 25 19.31(S)-(−)-Cyclohexylethylamine Hexetidine (mixture of isomers) Hydrogen 25(S)-(−)-Cyclohexylethylamine Undecylamine Hydrogen 25 10.88(S)-(−)-Cyclohexylethylamine Dehydroabietylamine Hydrogen 25(S)-(−)-2-Amino-3-phenyl-1- Hexetidine (mixture of isomers) Hydrogen 25propanol (S)-(−)-2-Amino-3-phenyl-1- Undecylamine Hydrogen 25 propanol(S)-(−)-2-Amino-3-phenyl-1- Dehydroabietylamine Hydrogen 25 propanol(1S,2S)-(+)-2-Amino-3- Hexetidine (mixture of isomers) Hydrogen 25methoxy-1-phenyl-1-propanol Octadecylamine (+)-Bornylamine Hydrogen 25Octadecylamine 1-Adamantanamine Hydrogen 25 Geranylamine2,3-Dimethylcyclohexylamine Hydrogen 25 14.53 Geranylaminetert-Octylamine Hydrogen 25 15.22 Geranylamine 1-AdamantanemethylamineHydrogen 25 4.37 Geranylamine Decahydroquinoline Hydrogen 25 31.79Geranylamine Dibenzylamine Hydrogen 25 6.48 GeranylamineN-Butylbenzylamine Hydrogen 25 16.44 Geranylamine CyclooctylamineHydrogen 25 12.37 Geranylamine (−)-Isopinocampheylamine Hydrogen 25 8.95Geranylamine 1-(1-Adamantyl)ethylamine, Hydrogen 25 32.95 HClGeranylamine Undecylamine Hydrogen 25 Geranylamine1-(1-Naphthyl)ethylamine Hydrogen 25 Amylamine 1-Adamantanamine Hydrogen37.5 0 3-Phenyl-1-propylamine 3,3-Diphenylpropylamine Hydrogen 37.53-Phenyl-1-propylamine 2,2-Diphenylamine Hydrogen 37.53-Phenyl-1-propylamine 1-Adamantanamine Hydrogen 37.5 18.652,2-Diphenylamine 3,3-Diphenylpropylamine Hydrogen 37.52,2-Diphenylamine 2,2-Diphenylamine Hydrogen 37.5 5.56 2,2-Diphenylamine1,3,3-Trimethyl-6- Hydrogen 37.5 8.67 azabicyclo[3.2.1]octane2,2-Diphenylamine 1-Adamantanamine Hydrogen 37.5 58.104-(Trifluoromethyl)benzylamine tert-Octylamine Hydrogen 37.5 7.474-(Trifluoromethyl)benzylamine 138 Hydrogen 37.5 4-Methylbenzylamine2-Fluorobenzylamine Hydrogen 50 22.10 4-Methylbenzylamine4-Fluorobenzylamine Hydrogen 50 14.62 4-Methylbenzylaminealpha-Methyltryptamine Hydrogen 50 0 4-Methylbenzylamine UndecylamineHydrogen 50 0 Cyclopentylamine Undecylamine Hydrogen 50 0 Furfurylamine2-Fluorobenzylamine Hydrogen 50 0 Furfurylamine Benzylamine Hydrogen 500 Furfurylamine 4-Fluorobenzylamine Hydrogen 50 0 Furfurylaminealpha-Methyltryptamine Hydrogen 50 0 Furfurylamine Undecylamine Hydrogen50 0 Furfurylamine Dehydroabietylamine Hydrogen 50 0 FurfurylamineFurfurylamine Hydrogen 50 0 3,4,5-Trimethoxybenzylamine2-Fluorobenzylamine Hydrogen 50 0 3,4,5-TrimethoxybenzylamineBenzylamine Hydrogen 50 0 3,4,5-Trimethoxybenzylaminealpha-Methyltryptamine Hydrogen 50 0 3,4,5-TrimethoxybenzylamineUndecylamine Hydrogen 50 0 3,4,5-TrimethoxybenzylamineDehydroabietylamine Hydrogen 50 0 1-Methyl-3-phenylpropylaminealpha-Methyltryptamine Hydrogen 50 0 1-Methyl-3-phenylpropylamineOctadecylamine Hydrogen 50 0 Cyclobutylamine Octadecylamine Hydrogen 500 Cyclobutylamine Undecylamine Hydrogen 50 0 CyclobutylamineDehydroabietylamine Hydrogen 50 0 1,2,3,4-Tetrahydro-1- Hexetidine(mixture of isomers) Hydrogen 50 0 naphthylamine 1,2,3,4-Tetrahydro-1-Aminodiphenylmethane Hydrogen 50 4.31 naphthylamine1,2,3,4-Tetrahydro-1- alpha-Methyltryptamine Hydrogen 50 0 naphthylamine1,2,3,4-Tetrahydro-1- 2-Methoxyphenethylamine Hydrogen 50 0naphthylamine 2,3-Dimethylcyclohexylamine Hexetidine (mixture ofisomers) Hydrogen 50 0 2,3-Dimethylcyclohexylamine AminodiphenylmethaneHydrogen 50 3.64 2,3-Dimethylcyclohexylamine alpha-MethyltryptamineHydrogen 50 0 Tyramine Furfurylamine Hydrogen 50 0 Tyramine2-Fluorobenzylamine Hydrogen 50 4.07 Tyramine Benzylamine Hydrogen 50 0Tyramine 2,4-Dichlorophenethylamine Hydrogen 50 0 2-FluorobenzylamineAminodiphenylmethane Hydrogen 50 0 2-Fluorobenzylamine4-Phenylbutylamine Hydrogen 50 0 2-Fluorobenzylamine2-Methoxyphenethylamine Hydrogen 50 0 2-Fluorobenzylamine2,4-Dichlorophenethylamine Hydrogen 50 0 2-Fluorobenzylamine1,3-Dimethylbutylamine Hydrogen 50 0 2-Fluorobenzylamine1-(1-Adamantyl)ethylamine, Hydrogen 50 0 HCl (R)-2-Amino-1-butanolDehydroabietylamine Hydrogen 50 0 3,4-DimethoxyphenethylamineAminodiphenylmethane Hydrogen 50 0 3,4-Dimethoxyphenethylamine4-Phenylbutylamine Hydrogen 50 0 3,4-Dimethoxyphenethylamine2-Methoxyphenethylamine Hydrogen 50 0 3,4-Dimethoxyphenethylamine2,4-Dichlorophenethylamine Hydrogen 50 0 3,4-Dimethoxyphenethylamine1,3-Dimethylbutylamine Hydrogen 50 0 3,3-Diphenylpropylamine PiperidineHydrogen 50 0 3,3-Diphenylpropylamine 2,3-Dimethylcyclohexylamine Methyl50 7.81 3,3-Diphenylpropylamine (−)-Isopinocamphenylamine Methyl 5013.06 Propylamine (S)-(+)-1-Amino-2-propanol Hydrogen 50 0Phenethylamine (S)-(+)-1-Amino-2-propanol Hydrogen 50 0 Phenethylamine4-Phenylbutylamine Hydrogen 50 0 Phenethylamine2,4-Dichlorophenethylamine Hydrogen 50 0 Phenethylamine1,3-Dimethylbutylamine Hydrogen 50 0 Phenethylamine1-(1-Adamantyl)ethylamine, Hydrogen 50 0 HCl Phenethylamine1-(1-Naphthyl)ethylamine Hydrogen 50 0 4-(2-Aminoethyl)morpholine2-Amino-2-methyl- 1-propanol Hydrogen 50 0 Cyclohexylamine2,4-Dichlorophenethylamine Hydrogen 50 0 exo-Aminonorbornane BenzylamineHydrogen 50 0 (+)-Isopinocampheylamine Hexetidine (mixture of isomers)Hydrogen 50 0 (+)-Isopinocampheylamine Aminodiphenylmethane Hydrogen 505.07 (+)-Isopinocampheylamine 4-Phenylbutylamine Hydrogen 50 0(+)-Isopinocampheylamine 2,4-Dichlorophenethylamine Hydrogen 50 0(+)-Isopinocampheylamine Undecylamine Hydrogen 50 0 Benzylamine3,3-Diphenylpropylamine Hydrogen 50 Benzylamine2-Amino-2-methyl-1-propanol Hydrogen 50 Benzylamine1-(1-Naphthyl)ethylamine Hydrogen 50 Benzylamine2,4-Dichlorophenethylamine Hydrogen 50 3-Amino-1-propanol UndecylamineHydrogen 50 0 2-Fluorophenethylamine 3,3-Diphenylpropylamine Hydrogen 500 2-Fluorophenethylamine 1-Adamantanemethylamine Hydrogen 50 02-Fluorophenethylamine 1-(3-Aminopropyl)-2- Hydrogen 50 0 pyrrolidinone(tech) 2-Fluorophenethylamine Decahydroquinoline Hydrogen 50 02-Fluorophenethylamine 1-Adamantanamine Hydrogen 50 24.342-Fluorophenethylamine 2,4-Dichlorophenethylamine Hydrogen 50 02-Fluorophenethylamine Undecylamine Hydrogen 50 0 2-FluorophenethylamineDehydroabietylamine Hydrogen 50 0 2-Fluorophenethylamine2-(1-Cyclohexenyl)ethylamine Methyl 50 0 2-FluorophenethylamineCyclooctylamine Methyl 50 5.81 b-Methylphenethylamine3,3-Diphenylpropylamine Hydrogen 50 0 b-Methylphenethylaminetert-Octylamine Hydrogen 50 0 b-Methylphenethylamine2-(1-Cyclohexenyl)ethylamine Hydrogen 50 0 b-Methylphenethylamine2-Amino-2-methyl-1-propanol Hydrogen 50 0 b-Methylphenethylamine4-Fluorophenethylamine Hydrogen 50 0 b-Methylphenethylamine GeranylamineHydrogen 50 0 b-Methylphenethylamine 5-Methoxytryptamine Hydrogen 50 04-Methoxyphenethylamine 3,3-Diphenylpropylamine Hydrogen 50 04-Methoxyphenethylamine 2-Amino-2-methyl-1-propanol Hydrogen 50 04-Methoxyphenethylamine 2,4-Dichlorophenethylamine Hydrogen 50 04-Methoxyphenethylamine 1-(1-Naphthyl)ethylamine Hydrogen 50 0L-Methioninol Hexetidine (mixture of isomers) Hydrogen 50 0Tetrahydrofurfuryl amine 1-Adamantanemethylamine Hydrogen 50 0Tetrahydrofurfurylamine 2-(1-Cyclohexenyl)ethylamine Hydrogen 50 0Tetrahydrofurfurylamine 4-Fluorophenethylamine Hydrogen 50 0Tetrahydrofurfurylamine Undecylamine Hydrogen 50 0 Amylamine1-Adamantanemethylamine Hydrogen 50 0 Amylamine Hexetidine (mixture ofisomers) Hydrogen 50 0 Amylamine Undecylamine Hydrogen 50 0 AmylamineDehydroabietylamine Hydrogen 50 0 1-Adamantanemethylaminecis-(−)-Myrtanylamine Methyl 50 0 3-Phenyl-1-propylamine4-(2-Aminoethyl)morpholine Hydrogen 50 3-Phenyl-1-propylamine1-(3-Aminopropyl)-2- Hydrogen 50 pyrrolidinone (tech)3-Phenyl-1-propylamine Veratryl amine Hydrogen 50 3-Phenyl-1-propylamine Aminodiphenylmethane Hydrogen 50 3-Phenyl-1-propylamineAminomethyl)phenylthio)benzyl Hydrogen 50 alcohol 2,2-Diphenylamine2-Fluorophenethylamine Hydrogen 50 2,2-Diphenylamine3,3-Diphenylpropylamine Methyl 50 2,2-Diphenylamine(+)-Isopinocampheylamine Methyl 50 2,2-Diphenylamine (+)-BornylamineMethyl 50 2,2-Diphenylamine Cyclooctylamine Methyl 50 2,2-Diphenylamine(−)-Isopinocampheylamine Methyl 50 3.81 4-(Trifluoromethyl)benzylamine4-(2-Aminoethyl)morpholine Hydrogen 50 4-(Trifluoromethyl)benzylamine2-(1-Cyclohexenyl)ethylamine Hydrogen 50 4-(Trifluoromethyl)benzylamineHexetidine (mixture of isomers) Hydrogen 504-(Trifluoromethyl)benzylamine 2,4-Dichlorophenethylamine Hydrogen 504-(Trifluoromethyl)benzylamine (S)-(−)-Cyclohexylethylamine Hydrogen 50Veratryl amine 1-Adamantanemethylamine Hydrogen 50 Veratryl amine2-(1-Cyclohexenyl)ethylamine Hydrogen 50 Veratryl amine4-Fluorophenethylamine Hydrogen 50 Veratryl amine Hexetidine (mixture ofisomers) Hydrogen 50 Veratryl amine 2,4-Dichlorophenethylamine Hydrogen50 Veratryl amine (S)-(−)-Cyclohexylethylamine Hydrogen 50 Veratrylamine Undecylamine Hydrogen 50 Veratryl amine DehydroabietylamineHydrogen 50 Veratryl amine 1-(1-Naphthyl)ethylamine Hydrogen 505-Amino-1-pentanol 1-Adamantanemethylamine Hydrogen 505-Amino-1-pentanol Dibenzylamine Hydrogen 50 5-Amino-1-pentanolcis-(−)-Myrtanylamine Hydrogen 50 12.97 2-(1-Cyclohexenyl)ethylamine2,4-Dimethoxybenzylamine Hydrogen 50 1-Aminomethyl-1- tert-AmylamineHydrogen 50 cyclohexanol, HCl 1-Aminomethyl-1-2-(2-Aminomethyl)phenylthio)benzyl- Hydrogen 50 cyclohexanol, HClalcohol 1-Aminomethyl-1- Undecylamine Hydrogen 50 cyclohexanol, HCl1-Aminomethyl-1- 1-(1-Naphthyl)ethylamine Hydrogen 50 cyclohexanol, HCl3-Fluorobenzylamine tert-Amylamine Hydrogen 50 3-FluorobenzylamineHexetidine (mixture of isomers) Hydrogen 50 3-FluorobenzylamineUndecylamine Hydrogen 50 4-Amino-1-butanol Undecylamine Hydrogen 504-Amino-1-butanol Dehydroabietylamine Hydrogen 502,4-Dimethoxybenzylamine N-Phenylethyldiamine Hydrogen 502,4-Dimethoxybenzylamine Aminodiphenylmethane Hydrogen 502,4-Dimethoxybenzylamine 4-Phenylbutylamine Hydrogen 502,4-Dimethoxybenzylamine 2-Chlorobenzylamine Hydrogen 502,4-Dimethoxybenzylamine 2,4-Dichlorophenethylamine Hydrogen 502,4-Dimethoxybenzylamine 2-(2-Chlorophenyl)ethylamine Hydrogen 502,4-Dimethoxybenzylamine 4-(Trifluoromethoxy)benzylamine Hydrogen 502-Ethoxybenzylamine Aminodiphenylmethane Hydrogen 50 2-Ethoxybenzylamine4-Phenylbutylamine Hydrogen 50 2-Ethoxybenzylamine 2-ChlorobenzylamineHydrogen 50 2-Ethoxybenzylamine 2-Aminoindan, HCl Hydrogen 502-Ethoxybenzylamine 2,5-Dimethoxyphenethylamine Hydrogen 502-Ethoxybenzylamine 4-(Trifluoromethoxy)benzylamine Hydrogen 502-Ethoxybenzylamine 1-(1-Naphthyl)ethylamine Hydrogen 50cis-(−)-Myrtanylamine 4-(2-Aminoethyl)morpholine Hydrogen 50cis-(−)-Myrtanylamine 2-Fluorophenethylamine Hydrogen 50cis-(−)-Myrtanylamine 1-(3-Aminopropyl)-2- Hydrogen 50 pyrrolidinone(tech) cis-(−)-Myrtanylamine Veratryl amine Hydrogen 50cis-(−)-Myrtanylamine N-Butylbenzylamine Hydrogen 50cis-(−)-Myrtanylamine 2,4-Dimethoxybenzylamine Hydrogen 50cis-(−)-Myrtanylamine 1,2,3,4-Tetrahydropyridoindole Hydrogen 50cis-(−)-Myrtanylamine 4-Phenylbutylamine Hydrogen 50cis-(−)-Myrtanylamine 2-(2-Chlorophenyl)ethylamine Hydrogen 50 3.91cis-(−)-Myrtanylamine 1-(1-Adamantyl)ethylamine, Hydrogen 50 10.85 HClcis-(−)-Myrtanylamine (R)-(−)-Cyclohexylethylamine Hydrogen 50 5.89cis-(−)-Myrtanylamine Dehydroabietylamine Hydrogen 50cis-(−)-Myrtanylamine 1-(1-Naphthyl)ethylamine Hydrogen 50cis-(−)-Myrtanylamine (+)-Bornylamine Methyl 50 4.04 Cyclooctylamine4-Methylcyclohexylamine Hydrogen 50 4.55 CyclooctylamineN-Phenylethyldiamine Hydrogen 50 Cyclooctylamine4-(Hexacylamino)benzylamine Hydrogen 50 Cyclooctylamine2,5-Dimethoxyphenethylamine Hydrogen 50 Cyclooctylamine2,4-Dichlorophenethylamine Hydrogen 50 3.36 Cyclooctylamine2-(2-Chlorophenyl)ethylamine Hydrogen 50 9.15 Cyclooctylamine1-(1-Adamantyl)ethylamine, Hydrogen 50 10.62 HCl Cyclooctylamine(S)-(−)-Cyclohexylethylamine Hydrogen 50 5.85 Cyclooctylamine(R)-(−)-Cyclohexylethylamine Hydrogen 50 Cyclooctylamine4-(Trifluoromethoxy)benzylamine Hydrogen 50 4.54 2-Adamantanamine, HClcis-(−)-Myrtanylamine Hydrogen 50 49.73 4-MethylcyclohexylamineN-Phenylethyldiamine Hydrogen 50 4-Methylcyclohexylamine4-Phenylbutylamine Hydrogen 50 4-FluorobenzylamineN-Benzyl-2-phenethylamine Hydrogen 50 4-Fluorobenzylamine Hexetidine(mixture of isomers) Hydrogen 50 4-Fluorobenzylamine UndecylamineHydrogen 50 4-Fluorobenzylamine Dehydroabietylamine Hydrogen 50 trans-2-Hexetidine (mixture of isomers) Hydrogen 50 Phenylcyclopropylamine, HCltrans-2- Undecylamine Hydrogen 50 Phenylcyclopropylamine, HCl trans-2-Dehydroabietylamine Hydrogen 50 Phenylcyclopropylamine, HCl(R)-1-Amino-2-propanol 4-(Hexacylamino)benzylamine Hydrogen 50(R)-1-Amino-2-propanol Undecylamine Hydrogen 50 (R)-I-Amino-2-propanolDehydroabietylamine Hydrogen 50 I-Leucinol Undecylamine Hydrogen 50(−)-Isopinocampheylamine 2-Ethoxybenzylamine Hydrogen 50 27.27(−)-Isopinocampheylamine Hexetidine (mixture of isomers) Hydrogen 50(−)-Isopinocampheylamine 4-Phenylbutylamine Hydrogen 50(−)-Isopinocampheylamine Dehydroabietylamine Hydrogen 50(−)-Isopinocampheylamine 1-(1-Naphthyl)ethylamine Hydrogen 50 Allylamine3,3-Diphenylpropylamine Hydrogen 50 Allylamine 2-Amino-1-propanol, d,lHydrogen 50 Allylamine Undecylamine Hydrogen 50 3-Amino-1,2-propanediolDehydroabietylamine Hydrogen 50 3-Ethoxypropylamine 2,2-DiphenylamineHydrogen 50 95.81 3-Ethoxypropylamine cis-(−)-Myrtanylamine Hydrogen 502-Aminoheptane Aminomethyl)phenylthio)benzyl Hydrogen 50 alcohol1-Naphthalenemethylamine Geranylamine Hydrogen 501-Naphthalenemethylamine Dehydroabietylamine Hydrogen 501-Aminopyrrolidine, HCl Hexetidine (mixture of isomers) Hydrogen 501-Aminopyrrolidine, HCl Undecylamine Hydrogen 50 1-Aminopyrrolidine, HClDehydroabietylamine Hydrogen 50 Ethanolamine 3,3-DiphenylpropylamineHydrogen 50 3-Methylbenzylamine Geranylamine Hydrogen 503-Methylbenzylamine 5-Methoxytryptamine Hydrogen 50 PiperonylamineAminodiphenylmethane Hydrogen 50 Piperonylamine2,4-Dichlorophenethylamine Hydrogen 50 Piperonylamine2-(2-Aminomethyl)phenylthio)benzyl Hydrogen 50 alcohol IsopropylamineDehydroabietylamine Hydrogen 50 4-Fluorophenethylamine2,4-Dimethoxybenzylamine Hydrogen 50 4-FluorophenethylamineAminodiphenylmethane Hydrogen 50 4-Fluorophenethylamine2-(2-Aminomethyl)phenylthio)benzyl Hydrogen 50 alcohol4-Chloroamphetamine, HCl N-Allylcyclopentylamine Hydrogen 50 10.254-Chloroamphetamine, HCl Hexetidine (mixture of isomers) Hydrogen 504-Chloroamphetamine, HCl 4-Phenylbutylamine Hydrogen 504-Chloroamphetamine, HCl 2-Methoxyphenethylamine Hydrogen 504-Chloroamphetamine, HCl Undecylamine Hydrogen 50 4-Chloroamphetamine,HCl Dehydroabietylamine Hydrogen 50 3-Fluorophenethylamine(−)-Isopinocampheylamine Hydrogen 50 3-Fluorophenethylamine1-Adamantamine Hydrogen 50 8.59 3-Fluorophenethylamine4-Phenylbutylamine Hydrogen 50 2-Methylcyclohexylamine (mix UndecylamineHydrogen 50 of cis and trans) 2-Methoxyphenethylamine3,3-Diphenylpropylamine Hydrogen 50 2-Methoxyphenethylamine(+)-Bornylamine Hydrogen 50 2-Methoxyphenethylamine tert-OctylamineHydrogen 50 20.46 2-Methoxyphenethylamine 1-AdamantanemethylamineHydrogen 50 2-Methoxyphenethylamine Dibenzylamine Hydrogen 502-Methoxyphenethylamine N-Butylbenzylamine Hydrogen 50 5.202-Methoxyphenethylamine 1,3,3-Trimethyl-6- Hydrogen 50 8.59azabicyclo[3.2.1]octane 2-Methoxyphenethylamine N-PhenylethyldiamineHydrogen 50 2-Methoxyphenethylamine 2,4-Dichlorophenethylamine Hydrogen50 2-Methoxyphenethylamine 2-(2-Chlorophenyl)ethylamine Hydrogen 502-Methoxyphenethylamine 1-(1-Adamantyl)ethylamine, Hydrogen 50 3.61 HCl2-Aminoindan, HCl (+)-Bornylamine Hydrogen 50 2-Aminoindan, HClNoradamantamine, HCl Hydrogen 50 7.43 2-(2-Chlorophenyl)ethylamineN-Phenylethyldiamine Hydrogen 50 2-(2-Chlorophenyl)ethylamineAminodiphenylmethane Hydrogen 50 2-(2-Chlorophenyl)ethylamine2,4-Dichlorophenethylamine Hydrogen 50 2-(2-Chlorophenyl)ethylamine1-(1-Adamantyl)ethylamine, Hydrogen 50 HCl 2-(2-Chlorophenyl)ethylamineDehydroabietylamine Hydrogen 50 2-(2-Aminomethyl)phenylthio)benzyl2-Methoxyphenethylamine Hydrogen 50 alcohol2-(2-Aminomethyl)phenylthio)benzyl 2,5-Dimethoxyphenethylamine Hydrogen50 alcohol 2-(2-Aminomethyl)phenylthio)benzyl2-(2-Chlorophenyl)ethylamine Hydrogen 50 alcohol2-(2-Aminomethyl)phenylthio)benzyl 1-(1-Adamantyl)ethylamine, Hydrogen50 alcohol HCl 2-(2-Aminomethyl)phenylthio)benzyl DehydroabietylamineHydrogen 50 alcohol 1-Aminoindan 4-Phenylbutylamine Hydrogen 501-Aminoindan 2,4-Dichlorophenethylamine Hydrogen 501,3-Dimethylbutylamine 4-Phenylbutylamine Hydrogen 50(S)-(−)-Cyclohexylethylamine Aminodiphenylmethane Hydrogen 50(S)-(−)-Cyclohexylethylamine 4-Phenylbutylamine Hydrogen 50(S)-(−)-Cyclohexylethylamine 2,4-Dichlorophenethylamine Hydrogen 50(S)-(−)-Cyclohexylethylamine 1-(1-Adamantyl)ethylamine, Hydrogen 50 HCl(1S,2S)-(+)-2-Amino-3- Dehydroabietylamine Hydrogen 50methoxy-1-phenyl-1-propanol Octadecylamine 2-Adamantanamine, HClHydrogen 50 3-Hydroxytyramine (1R,2S)-(−)-2-Amino-1,2- Hydrogen 50diphenylethanol 3-Hydroxytyramine Dehydroabietylamine Hydrogen 50Geranylamine 3,3-Diphenylpropylamine Hydrogen 50 GeranylamineN-Phenylethyldiamine Hydrogen 50 Geranylamine Hexetidine (mixture ofisomers) Hydrogen 50 Geranylamine 2-Thiopheneethylamine Hydrogen 50Geranylamine 2-Methoxyphenethylamine Hydrogen 50 Geranylamine2,5-Dimethoxyphenethylamine Hydrogen 50 Geranylamine2,4-Dichlorophenethylamine Hydrogen 50 Geranylamine2-(2-Chlorophenyl)ethylamine Hydrogen 50 2-Fluorophenethylamine2,3-Dimethylcyclohexylamine Methyl >50 2.074-(Trifluoromethyl)benzylamine 2,3-Dimethylcyclohexylamine Hydrogen >508.20 4-(Trifluoromethyl)benzylamine 1-Adamantanamine Hydrogen >50 32.025-Aminoquinoline exo-Aminonorbornane Hydrogen >50 17.87

[0138] TABLE 3 Compounds Synthesized in Larger Quantities for Further invitro Evaluations Cmpd Amount, Yields, # Name Structure mg % 1N-(4-Methylphenyl)-N′- (furfuryl)ethane-1,2-diamine

23 25 2 N-(4-Methylphenyl)-N′- (benzyl)ethane-1,2-diamine

27 29 3 N-[1-(1,2,3,4-Tetrahydro- naphthalene)-N′-(undecenyl)-ethane-1,2-diamine

11 10 4 N-[2-(3,4-Dimethoxy-phenyl)- ethyl-N′-(1-methyladamantyl)-ethane-1,2-diamine

13 11 5 N-[2-(3,4-Dimethoxy-phenyl)ethyl-N′-(norbornyl)-ethane-1,2-diamine

9 8 6 N-(1-Adamantylmethyl)-N′- (3,3-diphenylpropyl)propane- 1,2-diamine

55 36 7 N-(1-Adamantylmethyl)-N′- (3,3-diphenylpropyl)ethane-1,2-diamine

28 22 8 N-[2-(Cyclohexen-1 -yl)ethyl]- N-(3,3-diphenylpropyl)-propane-1,2-diamine

46 37 10 N-(−)-cis-Myrtanyl-N′-(3,3- diphenylpropyl)ethane-1,2- diamine

14 11 11 N-Cyclooctyl-N′-(3,3- diphenylpropyl)ethane-1,2- diamine

22 18 13 N-Allyl-N-cyclopentyl-N-(3,3- diphenylpropyl)ethane-1,2-diamine

33 27 14 N-(3,3-Diphenylpropyl)-N′- exo-(2-norborny)ethane-1,2- diamine

17 16 15 1-{2-[N-(3,3-Diphenylpropyl)]- aminoethyl}-3,5-dimethyl-piperidine

6.2 5 17 N-2-(2-Methoxyphenyl)ethyl- N′-(3,3-diphenylethyl)ethane-1,2-diamine

50 40 21 N-(3,3-Diphenylpropyl)-N′- (1S)-(1-ethylcyclohexane)-ethane-1,2-diamine

5 4 22 N-(3,3-Diphenylpropyl)-N′- (1R)-(1-ethylcyclohexane)-ethane-1,2-diamine

21 17 23 N-Allyl-N-cyclohexyl-N-(3,3- diphenylpropyl)ethane-1,2- diamine

6 5 24 N-2-(2-Methoxyphenyl)ethyl- N′-(4-fluorophenylethyl)-ethane-1,2-diamine

10 9 27 N-(3-Phenylpropyl)-N′-(1- adamantyl)ethane-1,2-diamine

11 10 28 N-(3-Phenylpropyl)-N′-(4- fluorophenyl)ethane-1,2- diamine

11 10 29 N-(2,2-Diphenylethyl)-N′-(2,3- dimethylcylcohexyl)ethane-1,2-diamine

4.5 4 31 N-(2,2-Diphenylethyl)-N′-(1S)- (1-ethylcyclohexane)-ethane-1,2-diamine

24 20 32 N-(2,2-Diphenylethyl)-N′-(R)-(+)-

58 48 33 N-(2,2-Diphenylethyl)-N′- (1,1,3,3-tetramethylbutyl)-ethane-1,2-diamine

11 9 34 N-(2,2-Diphenylethyl)-N′-(1- methyladamantyl)ethane-1,2- diamine

6.8 6 35 N-(2,2-Diphenylethyl)-N′- {1,1,3-trimethyl-6-azabicyclo-[3.2.1]octyl}ethane-1,2- diamine

38 20 36 N-{2-[N′-(2,2-Diphenylethyl)]- aminoethyl}-decahydroquinoline

28 24 37 N-(2,2-Diphenylethyl)-N′-(−)- cis-(myrtanyl)ethane-1,2- diamine

54 38 38 N-(−)-cis-(Myrtanyl)-N′-(2,2- diphenylethyl)propyl-1,2- diamine

39 30 40 N-(2,2-Diphenylethyl)-N′-(1R, 2R,3R,5S)-(−)-isopinocam-pheylethane-1,2-diamine

33 23 41 N-(−)-cis-(Myrtanyl)-N′-(2,3- dimethytcyclohexyl)ethane-1,2-diamine

66 62 42 N-(3,3-Diphenypropyl)-N′-(−)- cis-myrtanylethane-1,2- diamine

11 9 43 N-(−)-cis-Myrtanyl-N′-(1S,2S, 3S,5R)-(+)-isopinocampheylethane-1,2- diamine

31 27 47 N-(−)-cis-Myrtanyl-N′-(1R,2R, 3R,5S)-(−)-isopinocampheylethane-1,2- diamine

42 33 51 N-(Cyclooctyl)-N′-(2,3- dimethylcyclohexyl)ethane- 1,2-diamine

5.1 2 52 N-(Cyclooctyl)-N′-(3,3- diphenylpropyl)ethane-1,2- diamine

20 18 53 N-Cyclooctyl-N′-(1S,2S,3S, 5R)-(+)-isopinocampheyl-ethane-1,2-diamine

7.4 7 54 N-Cyclooctyl-N′-(R)-(+)- bornylethane-1,2-diamine

17 16 55 N-(Cyclooctyl)-N′-(1- methyladamantyl)ethane-1,2- diamine

7 6 56 N-(Cyclooctyl)-N′-(2S)-[2-(1- hydroxybutyl)]ethane-1,2- diamine

1.1 1 57 N-(−)-cis-Myrtanyl-N′- (cyclooctyl)ethane-1,2- diamine

18 18 58 N-(Cyclooctyl)-N′-(2- adamantyl)ethane-1,2- diamine

25 23 59 N-(Cyclooctyl)-N′-(1R,2R, 3R,5S)-(−)-isopinocampheylethane-1,2- diamine

15 14 61 N-(Cyclooctyl)-N′-[1-ethyl-(1- naphthyl)]ethane-1,2-diamine

16 14 62 N-(−)-cis-Myrtanyl-N′-(1S)-(1- ethylcyclohexane)ethane-1,2-diamine

48 46 63 N-(Cyclooctyl)-N′-trans-(2- phenylcyclopropyl)ethane-1,2-diamine

47 46 64 N-(2-Adamantyl)-N′-trans-(2- phenylcyclopropyl)ethane-1,2-diamine

49 46 65 N-(1-Adamantyl)-N′-trans-(2- phenylcyclopropyl)ethane-1,2-diamine

18 16 66 N-(3,3-Diphenylpropyl)-N′- (1R,2R,3R,5S)-(−)-isopinocampheylethane-1,2- diamine

2.3 2 68 N-(+/−)-[2-(1-Hydroxybutyl)]- N′-(1R,2R,3R,5S)-(−)-isopinocampheylethane-1,2- diamine

0.8 1 71 N-(1,1-Diphenylmethyl)-N′- (1R,2R,3R,58)-(−)-isopinocampheylethane-1,2- diamine

2.9 2 73 N-(2-Adamantyl)-N′-[2-(2- methoxyphenyl)ethyl]ethane-1,2-diamine

21 19 76 N-Allyl-N-cyclopentyl-N′-[2-(2- methoxyphenyl)ethyl]ethane-1,2-diamine

8 7 77 N-(1,1-Diphenylmethyl)-N′-[2- (2-methoxyphenyl)-ethyl]ethane-1,2-diamine

32 27 78 N-2-Adamantyl-N′-2,3- dihydro-1H-inden-2-yl- ethane-1,2-diamine

4.3 3 79 N-[2-(2,5-Dimethoxyphenyl)- ethyl]-N′-(R)-(+)-bornylethane-1,2-diamine

59 49 103 N,N′-Bis(cyclooctyl)ethane- 1,2-diamine

6.3 4 107 N-(2,2-Diphenylethyl)-N-(3- ethoxypropyl)ethane-1,2- diamine

58 52 109 N-Geranyl-N′-(2- adamanthyl)ethane-1,2-diamine

27 24 111 N-[2-(N′-Geranyl)aminoethyl]- 2-ethylpiperidine

24 24 116 N-Geranyl-N′-allyl-N′- (cyclopentyl)ethane-1,2- diamine

45 42 117 N-Geranyl-N′-(1,1-diphenyl- methyl)ethane-1,2-diamine

24 20 118 N-2-(2-Chlorophenyl)ethyl-N′- allyl-N′-(cyclopentyl)ethane-1,2-diamine

6.4 6 119 N-2-(2-Chlorophenyl)ethyl- N′-[2-(3-fluorophenyl)-ethyl]ethane-1,2-diamine

30 27 125 N,N′-bis-(−)-cis- Myrtanylpropane-1,2-diamine

41 35 134 N-[2-(N′-2,2-Diphenylethyl)- aminoethyll-(−)-3,4-dihydroxynorephedrine

20 15 151 N-[2-(2-Methoxy)phenylethyl]- N′-(1R,2R,3R,5S)-(−)-isopinocampheyl-ethane-1,2- diamine

67 60 164 N¹-[2-(4-fluorophenyl)ethyl]- N²-[2-(4-Methoxy)phenylethyl)-1-phenylethane- 1,2-diamine

94 73 165 N1-[2-(4-fluorophenyl)ethyl]- N2-(3-Phenylpropyl)-1-phenylethane-1,2-diamine

23 19

[0139] The present invention is also directed to a new library ofdiamine compounds useful against infectious disease. To further enhancethe structural diversity of prior diamine compounds, a synthetic schemeto incorporate amino acids into a bridging linker between the two aminecomponents has been developed. The use of amino acids allowed fordiverse linker elements, as well as chirality see FIG. 42 forrepresentative examples. The diamine compounds were prepared on mmolscale in 96-well format in pools of 10 compounds per well (for the vastmajority of the plates). Table 25 (FIG. 43) summarizes data for thesynthesized plates.

[0140] The reaction scheme followed is shown in FIG. 44.

[0141] Solid phase syntheses using Rink resin. Twenty one 96-well plateshave been prepared. Six-step synthetic route starting from the Rinkresin similar to what that had been used to create our first 100,000compound library (Scheme 1, FIG. 41), was applied to make targeteddiamines (Scheme 5, FIG. 44). Overall, all steps of these schemes aresimilar, except one (step 4) when formation of the second aminofunctionality occurs. In Scheme 1, the second amine is introduced intothe molecule as a whole synthon via nucleophilic displacement ofCl-function of the linker, while in the Scheme 5, it proceeds throughmodification of the existing amino moiety by carbonyl compounds.

[0142] Attachment of the first amine to the support was done accordingto the Garigipati protocol. Rink acid resin (Novabiochem) was convertedinto the Rink-chloride upon treatment with triphenylphosphine anddichloroethane in THF. This activated resin was then loaded by additionof an amine N1 in presence of Hunig's base in dichloroethane. The amineN1 includes, but is not limited to, alkyl and aryl primary amines. Outof 177 primary amines that had been previously used as N1 for 100,000library preparation, only 30 were selected in this Scheme, based upon invitro activity data of their ethylenediamine derivatives (from theprevious ˜100K library) as well as structural diversity (FIGS. 45 and46).

[0143] On the next step, the acylation reaction was accomplished viapeptide coupling with FMOC protected amino acids in presence of HATU(O-(7-Azabenzotriazol-1-yl)-N,N,N,N-tetramethyluroniumhexafluorophosphate) and EtN(iso-Pr)₂ in DCM/DMF mixture at roomtemperature. The reaction was done twice to improve product yields. Thelist of the amino acids used to create this library is shown in theTable 26 (FIG. 47).

[0144] Deprotection (removal of the FMOC group) was carried out byreaction with piperidine at room temperature. Derivatization of theamino group was achieved by reductive alkylation with various carbonylcompounds, such as aldehydes, ketones, and carboxylic acids, in thepresence of NaBCNH₃ at room temperature for 72-96 h. The selection ofthe carbonyl compounds was made so that the final diamine products wouldcarry the same or similar types of substituents that had been observedin the hit compounds generated from the previous library of ethambutolanalogs, as well as structural diversity (FIG. 48). A complete list ofthe carbonyl compounds used is shown in Table 27 (FIG. 49).

[0145] Reduction of the aminoethyleneamides into corresponding diamineswas carried out using the soluble reducing reagent 65+w % Red-A1 at roomtemperature. Cleavage of the products from the resin was achieved with a10% solution of trifluoroacetic acid in dichloromethane resulting in theformation of TFA salts of the diamines.

[0146] For library production the first three steps of the syntheticscheme (resin activation, amine loading, and acylation) were carried outusing a Quest 210 Synthesizer on scale of 0.1-0.15 g of resin per tube.Following the acylation, formed resins were thoroughly washed, dried,and then groups of ten resins were pooled together. A small amount ofeach resin (˜0.05 g) was archived prior to pooling to facilitatere-synthesis and deconvolution of actives.

[0147] Deprotection of the FMOC group, addition of the carbonylcomponent, reduction, and cleavage were carried out in 96-well reactionblocks using the Combiclamps system by Whatman Polyfiltronics or theFlexChem system by Robbins Scientific. A suspension of the pooled resinsin 2:1 mixture of DCM/THF was evenly distributed into one reaction plateresulting in approximately 10 mg of the resin per well. The 96 diversecarbonyl compounds were arrayed in one 96-well plate template and added,one carbonyl compound per well, to each individual pool of ten resins,resulting in an anticipated 960 diamines produced per plate. Reductionwas carried out in the same format and cleavage and filtering intostorage plates was followed by evaporation of the TFA prior tobiological assay.

[0148] Quality assessment of the prepared compounds was done byElectrospray Ionization mass spectrometry using two randomly selectedrows (16 samples) per plate, 17% of the total number. Successfulproduction of a compound was based on an appearance of a molecular ionof the calculated mass. Depending on the amino acid that had been usedfor the synthesis, the percentage of the predicted ions were observed,and therefore the predicted compounds were formed, varied from 5-60%(Table 25, FIG. 43). Based on MS analysis, out of targeted 20,000compounds, 4,500 diamines were actually formed.

[0149] Formulations

[0150] Therapeutics, including compositions containing the substitutedethylene diamine compounds of the present invention, can be prepared inphysiologically acceptable formulations, such as in pharmaceuticallyacceptable carriers, using known techniques. For example, a substitutedethylene diamine compound is combined with a pharmaceutically acceptableexcipient to form a therapeutic composition.

[0151] The compositions of the present invention may be administered inthe form of a solid, liquid or aerosol. Examples of solid compositionsinclude pills, creams, soaps and implantable dosage units. Pills may beadministered orally. Therapeutic creams and anti-mycobacteria soaps maybe administered topically. Implantable dosage units may be administeredlocally, for example, in the lungs, or may be implanted for systematicrelease of the therapeutic composition, for example, subcutaneously.Examples of liquid compositions include formulations adapted forinjection intramuscularly, subcutaneously, intravenously,intraarterially, and formulations for topical and intraocularadministration. Examples of aerosol formulations include inhalerformulations for administration to the lungs.

[0152] A sustained release matrix, as used herein, is a matrix made ofmaterials, usually polymers, which are degradable by enzymatic oracid/base hydrolysis, or by dissolution. Once inserted into the body,the matrix is acted upon by enzymes and body fluids. The sustainedrelease matrix is chosen desirably from biocompatible materials,including, but not limited to, liposomes, polylactides, polyglycolide(polymer of glycolic acid), polylactide co-glycolide (coplymers oflactic acid and glycolic acid), polyanhydrides, poly(ortho)esters,polypeptides, hyaluronic acid, collagen, chondroitin sulfate, carboxylicacids, fatty acids, phospholipds, polysaccharides, nucleic acids,polyamino acids, amino acids such as phenylalanine, tyrosine,isoleucine, polynucleotides, polyvinyl propylene, polyvinylpyrrolidoneand silicone. A preferred biodegradable matrix is a matrix of one ofeither polylactide, polyglycolide, or polylactide co-glycolide.

[0153] The dosage of the composition will depend on the condition beingtreated, the particular composition used, and other clinical factors,such as weight and condition of the patient, and the route ofadministration. A suitable dosage may range from 100 to 0.1 mg/kg. Amore preferred dosage may range from 50 to 0.2 mg/kg. A more preferreddosage may range from 25 to 0.5 mg/kg. Tablets or other forms of mediamay contain from 1 to 1000 mg of the substituted ethylene diamine.Dosage ranges and schedules of administration similar to ethambutol orother anti-tuberculosis drugs may be used.

[0154] The composition may be administered in combination with othercompositions and procedures for the treatment of other disordersoccurring in combination with mycobacterial disease. For example,tuberculosis frequently occurs as a secondary complication associatedwith acquired immunodeficiency syndrome (AIDS). Patients undergoing AIDStreatment, which includes procedures such as surgery, radiation orchemotherapy, may benefit from the therapeutic methods and compositionsdescribed herein.

[0155] The following specific examples will illustrate the invention asit applies to the particular synthesis of the substituted ethylenediamine compounds, and the in vitro and in vivo suppression of thegrowth of colonies of M. tuberculosis. In additiona, the teachings of R.Lee et al. J. Comb. Chem 2003, 5, 172-187 are hereby incorporated byreference in their entirety. It will be appreciated that other examples,including minor variations in chemical procedures, will be apparent tothose skilled in the art, and that the invention is not limited to thesespecific illustrated examples.

EXAMPLE I Generating the Ethylene Diamine Library

[0156] The Rink-acid resin was obtained from NOVABIOCHEM® Inc., SanDiego, Calif. Solvents: acetonitrile, dichloromethane,dimethylformamide, ethylenedichloride, methanol and tetrahydrofuran werepurchased from ALDRICH®, Milwaukee, Wis., and used as received. Allother reagents were purchased from SIGMA-ALDRICH®, West Monroe Highland,Ill. Solid phase syntheses were performed on a QUEST® 210 Synthesizer,from ARGONAUT TECHNOLOGIES®, Foster City, Calif., with the aid ofcombinatorial chemistry equipment, from WHATMAN® POLYFILTRONICS® (Kent,England; Rockland, Mass.) and ROBBINS SCIENTIFIC®, Sunnyvale, Calif.Evaporation of solvents was done using SPEEDVAC® AES, from SAVANT®,Holbrook, N.Y. All necessary chromatographic separations were performedon a WATERS'ALLIANCE HT SYSTEM®, Milford, Mass. Analytical thin-layerchromatography was performed on MERCK® silica gel 60F₂₅₄ plates,purchased from SIGMA-ALDRICH®, West Monroe Highland, Ill.

[0157] The activation of the Rink-acid resin, the addition of the firstamine, and the acylation step were carried out in 10 ml tubes using theQUEST® 210 Synthesizer. The addition of the second amine, the reductionwith Red-AL, and the cleavage from the solid support were carried out in96-deep (2 ml) well, chemically resistant plates.

[0158] A. Activation of the Rink-Acid Resin

[0159] The Rink-acid resin had a coverage of 0.43-0.63 mmol of linkerper gram resin. Four to five grams of this resin were suspended in 80 mlof a 2:1 mixture of dichloromethane and tetrahydrofuran (THF), anddistributed into ten, 10 ml tubes, with 8 ml of resin suspension pertube. Each suspension was filtered and washed twice with THF. A solutionof triphenylphosphine (3.80 g, 14.5 mmol) in 30 ml of THF was prepared,and 3 ml of this solution was added to each tube, followed by theaddition of 3 ml of a solution of hexachloroethane in THF (3.39 g/14.3mmol hexachloroethane in 30 ml THF). After agitation for six hours atroom temperature, each activated resin was washed twice with THF andtwice with dichloromethane.

[0160] B. Addition of the First Amine

[0161] Each tube, containing the activated rink resin, was charged with3 ml of dichloroethane, 0.3 ml (1.74 mmol) N₁N-diisopropylethylamine(EtN(iPr)₂) and the corresponding amine (around 1 mmol). If the selectedamine was a solid at room temperature, it was added as a solution, or asuspension in DMF. Enough dichloroethane was added to each tube for afinal volume of 8 ml. The reaction mixture was heated at 45° C. for 6-8hours. The resins were filtered, washed with a 2:1 mixture ofdichloromethane and methanol (1×8 ml), then with methanol (2×8 ml), andthen dried under argon for 10 minutes.

[0162] C. Acylation with the Halo-Acylchloride

[0163] a. Acylation with Chloroacetyl Chloride. Each resin was prewashedwith THF (2×8 ml), and then charged with THF (8 ml), pyridine (0.3 ml,3.67 mmole) and chloroacetyl chloride (0.25 ml, 2.5 mmole). The reactionmixture was stirred for 8 hours at 45° C., and then for 6-8 hours atroom temperature. Each resin was filtered, washed with a 2:1 mixture ofdichloromethane/methanol (1×8 ml), methanol (2×8 ml) and THF (2×8 ml).The acylation was repeated using the same loading of reagents, but ashorter reaction time of 4 hours at 45° C., and 2 hours at roomtemperature. Each resin was then filtered, washed with a 2:1 mixture ofdichloromethane and methanol (1×8 ml), and then with methanol (3×8 ml).Each resin was dried under argon for 10 minutes. Each resin was thentransferred into a vial and dried in a desiccator under vacuum for 1hour.

[0164] b. Acylation with α-Phenyl-α-Chloroacetyl Chloride. The sameprocedure set out for the acylation with chloroacetyl chloride was used.A 2.5 mmol excess of α-phenyl-α-chloroacetyl chloride, relative to mmolamount of linker in the rink-acid resin, was used.

[0165] C. Acylation with α-Halo-α-Methyl; α-Halo-α-Ethyl andα-Halo-α-Butylacetyl Bromide. A 1:1:1 mixture (by volume) of theα-bromoproponyl bromide (R₄=Me), α-bromobutyryl bromide (R₄=Et), andα-bromohexanoyl bromide (R₄ Bu) was used to give a molar ratio of0.52:0.56:0.42 (in mmols). This resulted in a molar excess of 1.65, 1.75and 1.31, respectively, if the original coverage of the resin was 0.63mmol/g (0.5 g resin per tube), and 2.4, 2.6 and 1.9 if the originalcoverage of the resins was 0.43 mmol/g (0.5 g resin per tube).

[0166] d. Acylation with α-Chloro-α-Methyl Acetic acid. Each resin wasprewashed with dichloromethane. Each tube was charged with 3 ml of asolution of PyBrop (0.29 g, 0.62 mmole) in dichloromethane, a solutionof the α-chloro-α-methylacetic acid (0.095 g, 0.62 mmole) in 3 ml ofDMF, and EtN(iPr)₂ (0.2 ml, 1.2 mmole). Each reaction mixture wasallowed to react for 16-18 hours at room temperature. Each resin wasthen filtered, washed with dichcloromethane (2×8 ml) and methanol (2×8ml), and the acylation was repeated. Each resin was then filtered,washed with dichloromethane (2×8 ml), methanol (3×8 ml), and dried underargon for about 10 minutes. Each resin was transferred into a vial, anddried in a desiccator under vacuum for one hour.

[0167] D. Addition of the Second Amine

[0168] Ten, or thirty prepared α-haloacetyl amide resins from the firstthree steps were pooled together, leaving 0.05-0.10 gram of eachindividual resin for necessary deconvolutions. A suspension of thepooled resin mixture in 100 ml of a 2:1 mixture of dichloromethane andTHF was distributed into one, two or three, 96-well reaction plates. Forone reaction plate, 1.7 to 2.0 grams of resin were used. For tworeaction plates, 3.0 to 3.3 grams of-resin were used, and for threereaction plates, 4.7 to 5.0 grams of resin were used. The distributedsuspension was then filtered using a filtration manifold, a smalllightweight manifold that is generally used for drawing solvents andreagents from the chambers of the 96-well reaction plates. The reactionplates were transferred into COMBICLAMPS® (Huntington, W. Va.), and 10%EtN(iPr)₂ in DMF was added at 0.2 ml per well (0.21 mmole of EtN(iPr)₂per well), followed by the addition of a 1.0M solution of theappropriate amine from the corresponding master plate, 0.1 ml per well(0.1 mmole amine per well). The COMBICLAMPS® are used to accommodate96-well reaction plates during synthesis, allowing for the addition ofreagents into the plates, and a proper sealing that maintains reagentsand solvents for hours at elevated temperatures. These clamps consist ofa top and bottom cover provided with changeable, chemically resistantsealing gaskets. They are designed to accommodate 96-well reactionplates between the top and bottom covers. The reaction plates weresealed and kept in an oven at 70-75° C. for 16 hours. After cooling toroom temperature, the resins were filtered, washed with a 1:1 mixture ofDCM/methanol (1×1 ml), methanol (2×1 ml), and then dried in a desiccatorunder vacuum for 2 hours.

[0169] E. Reduction with Red-A1

[0170] The reaction plates were placed into COMBICLAMPS®. A 1:6 mixtureof Red-A1 (65+w % in toluene) and THF was added, at 0.6 ml per well(0.28 mmole of Red-Al per well), and allowed to react for 4 hours. Eachresin was then filtered, washed with THF (2×1 ml), and methanol (3×1ml). The addition of methanol should proceed with caution. Each resinwas then dried under vacuum.

[0171] F. Cleavage of Final Ethylene Diamine Compound

[0172] This step was carried out using a cleavage manifold, a Tefloncoated aluminum, filter/collection vacuum manifold, designed forrecovering cleavage products from the reaction plates into collectionplates. The manifold is designed to ensure that the filtrate from eachwell is directed to a corresponding well in a receiving 96-wellcollection plate. The reaction plates (placed on the top of thecollection plates in this manifold) were charged with a 10:85:5 mixtureof TFA, dichloromethane, and methanol (0.5 ml of mixture per well).After fifteen minutes, the solutions were filtered and collected intoproper wells on the collection plates. The procedure was repeated.Solvents were evaporated on a SPEED VAC®, Holbrook, N.Y., and theresidual samples (TFA salts) were tested without further purification.

EXAMPLE II Deconvolution Example

[0173] Deconvolution of the active wells was performed by re-synthesisof discrete compounds, from the archived α-haloacetyl amide resins (10resins, 0.05-0.10 g each), which were set aside at the end of theacylation step before the pooling. Each resin was assigned a discretecolumn (1, or 2, or 3, etc., see the template) in a 96 well filterplate,and was divided between X rows (A, B, C, etc), where X is the number ofhits discovered in the original screening plate. To each well, in a row,a selected N2 (R₃R₂NH) hit amine (0.1 mmol), DMF (180 ml) and EtNiPr₂(20 ml) were added: the first selected amine was added to the resins inthe row “A”, the second amine—to the resins in the row “B”, the thirdamine—to the resins in the row “C”, etc. A lay-out of a representative96-well filter plate is shown in Table 4.

[0174] The reaction plates were sealed and kept in an oven at 70-75° C.for 16 hours. After cooling to room temperature, the resins werefiltered, washed with a 1:1 mixture of DCM and methanol (1×1 ml),methanol (2×1 ml), and dried in desiccator under vacuum for 2 h.Reduction and cleavage were performed according to steps 5 and 6 in theoriginal synthetic protocol. The product wells from the cleavage wereanalyzed by ESI-MS (Electro Spray Ionization Mass Spectroscopy) toensure the identity of the actives, and were tested in the same Luc andMIC assays. TABLE 4 Lay-Out of Representative 96-Well Filter Plate A1 A2A3 A4 A5 A6 A7 A8 A9 A10 Selected amine N2, Added to A1-A10 B1 B2 B3 B4B5 B6 B7 B8 B9 B10 Selected amine N2, Added to B1-B10 C1 C2 C3 C4 C5 C6C7 C8 C9 C10 Selected amine N2, Added to C1-C10 D1 D2 D3 D4 D5 D6 D7 D8D9 D10 Selected amine N2, Added to D1-D10 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10Selected amine N2, Added to E1-E10 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10Selected amine N2, Added to F1-F10 G1 G2 G3 G4 G5 G6 G7 G8 G9 G10Selected amine N2, Added to G1-G10 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10Selected amine N2, Added to H1-H10 *X* selected Amines N2 to be added onthe step 4 Resin Resin Resin Resin Resin Resin Resin Resin Resin ResinIndividual resins ##1-10, #1 #2 #3 #4 #5 #6 #7 #8 #9 #10 preloaded withproper amine N1.

EXAMPLE III Solid-Phase Synthesis of Selected SubstitutedEthylenediamine Compounds Using the QUEST® 210 Synthesizer

[0175] The solid-phase protocol described above in Example I was appliedto the scaled-up synthesis of the selected substituted ethylene diaminecompounds. Here, all reaction steps, from the activation of theRink-acid resin to the cleavage of the final product, were carried outusing the QUEST® instrument only, which allowed for the simultaneoussyntheses of twenty parallel reactions. Purification of all crudesamples was done by HPLC to yield desirable products in purity greaterthan 90%. Table 3 lists the scale-ups of substituted ethylene diamines.Here, the synthesis of one of the active compounds,N-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine is described below as anexample.

[0176] The Preparation of N-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine(compound 109) is set forth in FIG. 12.

[0177] 1. Activation of the Rink-acid resin. Synthesis of Rink-Cl resin.Rink-acid resin, coverage (linker) of 0.43 to 0.63 mmol/g (0.8 g, 0.5mmol), was placed into one of the 10 ml tubes of QUEST® 210 Synthesizer,and washed twice with THF. A solution of triphenylphosphine (0.380 g,1.45 mmol) in THF (3 ml) was added, followed by the addition of asolution of hexachloroethane (0.4 g, 1.43 mmol) in THF (3 ml). THF wasadded up to the volume of the tube (approximately 2 ml). After 6 hours,the resin was filtered, washed with THF (2×8 ml) and dichloromethane(2×8 ml).

[0178] 2. Addition of the first amine. Synthesis of resin attachedgeranylamine. The tube with activated resin was charged with 3 ml ofdichloroethane, EtN(iPr)₂, (0.3 ml, 1.74 mmol), and geranylamine (0.230g, 1.5 mmol). Dichloroethane was added to a volume of 8 ml. The reactionwas carried for 8 hours at 45° C., and for 6-8 hours at roomtemperature. Geranylamine loaded resin was filtered, washed with a 2:1mixture of dichloromethane and methanol (1×8 ml), then with methanol(2×8 ml), and suck dried for 10 minutes under argon.

[0179] 3. Acylation with chloroacetyl chloride. Synthesis of resinattached N-Geranyl-α-chloroacetamide. The resin was prewashed with THF(2×8 ml). The tube was charged with 8 ml of THF, pyridine (0.3 ml, 3.67mmol), and chloroacetyl chloride (0.2 ml, 2.5 mmol), and allowed to stirfor 8 h at 45° C., and 6-8 h at room temperature (RT). After thereaction was complete, the resin was filtered, washed with a 2:1 mixtureof dichloromethane and methanol (1×8 ml), methanol (2×8 ml), and THF,and the acylation was repeated using the same loads of the reagents, butshorter reaction time: 4 hours at 45° C. and 2 hours at roomtemperature. At the end, the α-chloroacetamide loaded resin wasfiltered, washed with a 2:1 mixture of dichloromethane and methanol (1×8ml), methanol (3×8 ml), and suck dried for 15 min under argon.

[0180] 4. Addition of the second amine. Synthesis of resin attachedN-Geranyl-N′-(2-adamantyl)acetamide. The tube with the resin was chargedwith DMF (3 ml) and EtN(iPr)₂ (0.6 ml, 4.4 mmol), followed by theaddition of a suspension of 2-adamantamine hydrochloride (2.0 g, 1.1mmol) in DMF (4 ml), and was allowed to stir at 70-75° C. for 16 hours.After cooling down to the room temperature, the resin was filtered,washed with a 1:1 mixture of DCM and methanol (1×8 ml), methanol (2×8ml), and suck dried for 15 minutes under argon.

[0181] 5. Reduction with Red-A1. Synthesis of resin attachedN-Geranyl-N′-(2-adamantyl)ethane-1,2-diamine. The resultant resin wassuspended in anhydrous THF (3 ml) in a tube, and stirred for 15 min.Commercially available Red-A1, 65+w % in toluene, was added (2.0 ml, 6.4mmol), followed by addition of 2-3 ml of anhydrous THF (to fill up thevolume of the tube). The mixture was allowed to react for 4 hours. Afterthe reaction, the resin was filtered, washed with THF (1×8 ml), a 1:1mixture of THF and methanol (1×8 ml) (addition of MeOH should proceedwith caution), methanol (3×8 ml), and then dried.

[0182] 6. Cleavage from the resin and purification. Synthesis ofN-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine acetate. For this laststep of the synthesis, the tube with the resin was charged with a 10:90mixture of TFA and dichloromethane, and the formed bright red suspensionwas allowed to stir for 30 min. After addition of MeOH (0.5 ml), thecolorless suspension was filtered, and the filtrate was collected into aproper tube. The procedure was repeated, and solvents were evaporated ona SPEEDVAC®. Half of the amount of crudeN-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine (in a form oftrifluoroacetate salt) was purified by HPLC using following conditions:column C18, flow 4 ml/min, 30 min run, gradient starting with5%AcOH/MeOH (100%) finishing up with acetonitrile (100%). Obtained: 27mg of N-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine diacetate, 24%yield, 98% purity by NMR.

EXAMPLE IV Representative Solution Phase Synthesis of the ActiveCompounds

[0183] Preparation of N-(Cyclooctyl)-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheylethane-1,2-diamine as hydrochloride (compound 59)is set forth in FIG. 13.

[0184] Bromocyclooctylacetylamide. To a mixture of cyclooctylamine (3.3g, 0.026 mol) and pyridine (2.42 g, 0.031 mmol) in anhydrous THF (80 ml)at 0° C. was added dropwise, via syringe, bromoacetylbromide (5.78 g,0.029 mol). The reaction temperature was maintained by an ice bath. Thereaction mixture was allowed gradually to warm up to room temperature,and was stirred at room temperature for 1 hour. The precipitate wasremoved by filtration, washed with ethyl ether (1×30 ml), and thefiltrate was concentrated to dryness on a rotory evaporator.Bromocyclooctylacetylamide was forwarded to the second step withoutadditional purification.

[0185] N-(Cyclooctyl)-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheyl-1-carbonylethane-1,2-diamine. To a solution ofthe bromocyclooctylacetylamide in DMF (60 ml) were added Hunig's base(4.64 g, 0.036 mol) and (1R, 2R, 3R, 5S)-(−)-isopinocampheylamine (4.5g, 0.029 mol), and the reaction mixture was stirred at 80° C. for 16hours. After cooling off to the room temperature, the reaction mixturewas diluted with 150 ml of ethyl ether, and washed with 1M NaOH solution(2×50 ml). The organic layer was washed with brine (1×50 ml), dried overMgSO₄, and concentrated to dryness on the rotory evaporator. The residue(11.04 g) as brown oil was purified on COMBIFLASK® (Isco, Lincoln,Nebr., USA), using Silicagel catridges commercially available fromBIOTAGE® (Biotage, Inc. of Dyax Corp, Va., USA), and the followingmobile phase gradient: 30 min run, starting with DCM, 100%, andfinishing up with a mixture DCM:MeOH:NH₄OH (600:400:10). The finalproduct (7.29 g) was obtained as a brown oil; 76% yield, purity 90%.

[0186] N-(Cyclooctyl)-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheylethane-1,2-diamine. To a solution of the amide,from previous step, in anhydrous THF (160 ml), was added dropwise viasyringe commercially available (SIGMA-ALDRICH®) Red-A1, as 65 wt %solution in THF (28 ml, 0.09 mol). The reaction mixture was stirred atreflux for 20 hours. After cooling down to the room temperature, thereaction mixture was poured into 1.5M NaOH (200 ml), and extracted withethyl ether (2×100 ml). The organic layer was washed with brine (1×100ml), dried over MgSO₄, and evaporated to dryness on the rotoryevaporator to yield 7.2 g of a crude product, as a brown oil.Chromatographic purification of the crude using the same equipment andconditions as for the previous step, gave 3.5 g of the diamine. Thediamine was treated with 2.0M solution of HCl in ethyl ether (25 ml),and kept in a refrigerator overnight. A dark yellow solid (4.2 g)formed, and was filtered off, and recrystallized from MeOH and ethylether to yield 1.5 g of the diamine as an HCl salt (of purity greaterthan 98%, NMR and MS are available), 19% overall yield.

EXAMPLE V

[0187] Mass Spectroscopy Analysis

[0188] Mass spectra data were obtained by Elecrospray Ionizationtechnique on a PERKIN ELMER®/SCIEX®, API-300, TQMS with an autosampler,manufactured by SCIEX®, Toronto, Canada.

[0189] A. Library of Substituted Ethylenediamines

[0190] Mass spectroscopy served as a means for monitoring the reactionresults of the library of ethylenediamines. Mass spectroscopy was doneon two randomly selected rows (24 samples) per reaction plate, forroughly 28,000 compounds in pool of 10 or 30 compounds per well. Thus,if ten compounds per well were synthesized, the mass spectra for eachwell should contain ten signals, correlating with the proper molecularions for each compound. The presence or absence of a particular signalindicated the feasibility of the particular synthesis. Based on the massspectral data, and on a general analysis of the reactivity of thevarious amines, it is estimated that 67,000 compounds were formed out of112,000 compounds.

[0191]FIG. 14 is a representative mass spec profile for one sample well.Mass spectra for a representative ethylene diamine compound is shown inFIG. 15. Tables 5 to 8, below, list illustrative examples of mass specdata for representative reaction wells, with each well containing tensubstituted ethylene diamines. TABLE 5 ILLUSTRATIVE EXAMPLES OF MASSSPEC DATA FOR REPRESENTATIVE ETHYLENEDIAMINES (TEN COMPOUNDS PER WELL).R₂R₃NH in the 2^(nd) position (from the [M + 1)⁺of the R₁NH₂ in the1^(st) position master plate of the product (pool of 10 resins) amines)R₁NHCH₂CH₂NR₂R₃ Plate #4-034-2, well D10 1-(2-Aminoethyl)piperidine2-Aminoheptane 270 absent Phenethylamine 263 4-(2-Amino- 272 absentethyl)morpholine Tryptamine 302 Cyclohexylamine 241Exo-2-Aminonorbornane 253 Benzylamine 249 2-Fluorophenethylamine 281?-Methylphenethylamine 277 4-Methoxyphenethylamine 293 Plate #4-56-1,well C4 4-Methylbenzylamine exo-2- 259 Cyclopentylamine Aminobornane 2232-(Aminomethyl)piperidine 246 low intensity Furfurylamine 2353,4,5-Trimethoxybenzyl- 335 amine 1-Methyl-3-phenyl- 287 propylamineCylcobutylamine 209 1,2,3,4-Tetrahydro-1- 258 naphthylamine2,3-Dimethylcyclohexyl- 265 amine 2-Amino-1-butanol 227 low intensityPlate #4-44-2, well G1 Veratrylamine 4-Fluorophenethyl- 3332-(1-Cyclohexenyl)ethyl- amine 291 amine 5-Aminoquinolone 310 absent1-(1-Naphthyl)ethylamine 337 absent 1-Aminopiperidine 2663-Fluorobenzylamine 291 2,4-Dimethoxybenzylamine 3333-Amino-1,2,4-triazine 262 absent 2-Ethoxybenzylamine 317 4-(3-Amino-310 absent propyl)morpholine

[0192] TABLE 6 Mass Spec Data for Synthesized Ethylenediamines

R₁NH₂ in the [M + 1]⁺ of the [M + 1]⁺ of the products, R₄ = Ph 1^(st)position products, R₄ = H Diamines, 1 Amino alcohols, 13 Tyramine 308384 258 formed 2-Adamantamine 321 absent 398 absent 272 formedcis-Myrtanylamine 324 400 274 formed 3-Amino-1-propanol 246 322 196absent L-Methioninol 305 absent 382 absent 256 absent Cyclooctylamine298 374 248 formed (1S,2S)-2-Amino-1-phenyl- 337 absent 414 absent 288absent 1,3-propandiol 1-Adamantanemethylamine 336 412 absent 286 formed2,2-Diphenylethylamine 368 444 318 formed 5-Amino-1-pentanol 274 350 224formed

[0193] TABLE 7 Mass Spec Data for Synthesized Ethylenediamines, R₄ = Hand Me

R₁NH₂ in the [M + 1]⁺ of the [M + 1]⁺ of the products, R₄ = Ph 1^(st)position products, R₄ = H Diamines, 1 Amino alcohols, 13 Tyramine 278293 196 absent 2-Adamantamine 293 absent 307 absent 210 low intensitycis-Myrtanylamine 293 309 212 formed 3-Amino-1-propanol 217 231 134absent L-Methioninol 277 absent 291 absent 194 formed Cyclooctylamine269 269 absent 186 absent (1S,2S)-2-Amino-1-phenyl- 309 low intensity323 absent 226 formed 1,3-propandiol 1-Adamantanemethylamine 307 321 224formed 2,2-Diphenylethylamine 339 353 256 formed 5-Amino-1-pentanol 245259 162 absent

[0194] TABLE 8 Mass Spec Data for Synthesized Ethylenediamines, R₄ = Hand Me

R₁NH₂ in the [M + 1]⁺ of the [M + 1]⁺ of the products, R₄ = Ph 1^(st)position products, R₄ = H Diamines, 1 Amino alcohols, 13 Tyramine 278292 absent 196 absent 2-Adamantamine 292 absent 306 absent 210 formedcis-Myrtanylamine 294 308 absent 212 formed 3-Amino-1-propanol 216 230absent 134 absent L-Methioninol 276 absent 290 absent 194 absentCyclooctylamine 268 282 absent 186 absent (1S,2S)-2-Amino-1-phenyl- 308322 absent 226 formed 1,3-propandiol 1-Adamantanemethylamine 306 absent320 absent 224 formed 2,2-Diphenylethylamine 338 352 absent 256 formed5-Amino-1-pentanol 244 258 absent 162 absent

EXAMPLE VI ¹H NMR Spectroscopy

[0195] Proton NMR data was recorded on a VARIAN® Nuclear MagneticResonance Spectrometer (Palto Alto, Calif.) at 500 MHz.

[0196] Representative substituted ethylene diamines were purified byHPLC, and analyzed by proton NMR. A representative proton NMR profilesis shown in FIG. 16. NMR and MS data for some representative hitcompounds are shown below.

[0197] Compound 6.

[0198]N²-(1-Adamantylmethyl)-N¹-(3,3-diphenylpropyl)propane-1,2-diamine. 55mg, 36% yield. ¹H NMR: δ 7.28-7.15 (m, 5H), 3.95 (t, J=7.9 Hz, 1H), 2.94(br s 4H), 2.71 (dd, J=7.6, 9.8 Hz, 2H), 2.41 (s, 2H), 2.32 (dd, J=7.6,7.9 Hz, 2H), 2.16 (s), 2.08-1.98 (m, 4H), 1.72 (m, 6H), 1.62 (m, 6H),1.51 (d, J=2.4 Hz, 3H). Mass spectrum (ESI) m/z (MH)⁺417.

[0199] Compound 7

[0200] N-(3,3-Diphenylpropyl)-N′-(1-adamanthylmethyl)ethane-1,2-diamine.28 mg, 22% yield. ¹H NMR (500 MHz) δ 7.30-7.12 (m, 10H); 3.95 (t, J=7.6Hz, 1H); 2.91 (d, J=1.2 Hz, 4H); 2.70 (dd, J=7.6 and 1.2 Hz, 2H); 2.40(d, J=1.3 Hz, 2H); 2.32 (q, J=8.0 Hz, 2H); 1.98 (br d, J=1.7 Hz, 4H);1.72 (d, J=12.2 Hz, 4H); 1.62 (d, m? J=12.2 Hz, 4H); 1.51 (br s, 6H).Mass spectrum (ESI) m/z (MH)⁺403.6.

[0201] Compound 10.

[0202] N-(−)-cis-Myrtanyl-N′-(3,3-diphenylpropyl)ethane-1,2-diamine. 14mg, 11% yield. ¹H NMR (500 MHz) δ 7.30-7.10 (m, 10H); 3.95 (m, 1H);2.92-2.83 (m, 4H); AB: 2.80 (d, J=7 Hz, 1H); 2.76 (d, J=8 Hz, 1H); 2.65(dd, J=9.6 and 7.6 Hz, 2H); 2.42-2.20 (m, 4H), 2.29 (d, J=8 Hz, 2H),1.90 (m, 8H); 1.42 (m, 1H); 1.19 (m, 2H); 1.17 (s, 3H); 0.95 (s, 3H);1.00-0.8 (m, 2H). Mass spectrum (ESI) m/z (MH)⁺391.3.

[0203] Compound 14.

[0204] N-(3,3-Diphenylpropyl)-N′-exo-(2-norborny)ethane-1,2-diamine. 17mg, 16% yield. ¹H NMR (500 MHz) δ 7.30-7.15 (m, 10H); 3.95 (t, J=7.9 Hz,1H); 2.86 (dd, J=11.5 and 1.5 Hz, 4H); 2.73 (dd, J=8.0 and 3.3 Hz, 1H);2.64 (t, J=7.6 Hz, 2H); 2.29 (t, J=7.5 Hz, 2H), 2.31-2.26 (m, 2H) 2.301.96 (s, 3H); 1.63 (ddd, J=13.1, 7.9 and 2.5 Hz, 1H); 1.60-1.50 (m, 1H);1.50-1.43 (m, 2H); 1.30 (dq, J=4.0 and 13.5 Hz, 1H), (1H, m), 1.20 (dd,J=10.4 and 1.1 Hz, 1H), 1.11 (dd, J=2.0, and 8.5 Hz, 1H), 1.08 (dd,J=2.5, and 8.5 Hz, 1H), 1.10 (dq, J=8.3 and 2.1, 2H). Mass spectrum(ESI) m/z (MH)+349.1.

[0205] Compound 21.

[0206]N-(3,3-Diphenylpropyl)-N′-(1S)-(1-ethylcyclohexane)ethane-1,2-diamine. 5mg, 4% yield. Mass spectrum (ESI) m/z (MH)⁺365.5.

[0207] Compound 32.

[0208] N-(2,2-Diphenylethyl)-N′-®-(+)-bornylethane-1,2-diamine. 58 mg,48% yield. ¹H NMR (500 MHz): δ 7.30-7.10 (m, 10H); 4.18 (t, J=6.8 Hz,1H); 3.34 (d, J=7.6 Hz, 2H); 3.02 (m, 4H); 2.95-2.90 (m, 1H); 2.15-2.08(m, 1H); 1.94 (m, 1H); 1.72-1.65 (m, 2H); 1.48-1.30 (m, 2H); 1.27-1.10(m, 2H); 1.06 (dd, J=13.6 and 4.1 Hz, 1H); 0.82 (s, 3H); 0.81 (s, 3H);0.78 (s, 3H). Mass spectrum (ESI) m/z (MH)⁺377.2

[0209] Compound 34.

[0210] N-(2,2-Diphenylethyl)-N′-(1-adamanthylmethyl)ethane-1,2-diamine.6.8 mg, 6% yield. ¹H NMR (500 MHz) δ 7.30-7.15 □m, 10H); 4.15 (t, J=7.6Hz, 1H); 3.24 (dd, J=7.9 and 1.2 Hz, 2H); 2.79 (t, J=6.5 Hz, 2H); 2.74(t, J=6.0 Hz, m, 2H); 1.95 (m, 8H); 1.69 (d, J=12.5 Hz, 4H); 1.59 (d,J=11.9 Hz, 4H); 1.40 and 1.39 (br s, 3H); Mass spectrum (ESI) m/z(MH)⁺389.0.

[0211] Compound 37.

[0212] N-(2,2-Diphenylethyl)-N′-(−)-cis-myrtanylethane-1,2-diamine. 54mg, 38% yield. ¹H NMR: δ 7.31-7.18 (m, 10H), 4.13 (t, J=7.6 Hz, 1H),3.26 (d, J=7.6 Hz, 2H), 2.86 (dd, J=4.3, 8.0 Hz, 4H), 2.76 (dd, J=7.6,12.2 Hz, 2H), 2.37 (ddd, J=1.8, 9.0, 12.5 Hz, 1H), 2.12 (dq, J=1.8, 7.6Hz, 1H), 1.98 (br s, 2H), 1.98-1.84 (m, 4H), 1.39 (ddd, J=2.4, 4.0, 6.1Hz, 1H), 1.18 (s, 3H), 0.95 (s, 3H), 0.91 (d, J=10.0 Hz, 1H) Massspectrum (ESI) m/z (MH)⁺377.2.

[0213] Compound 38.

[0214] N-(−)-cis-Myrtanyl-N′-(2,2-diphenylethyl)propane-1,2-diamine. 39mg, 30% yield. ¹H NMR (500 MHz) δ 7.30-7.15 (m, 10H); 4.13 (t, J=8.0 Hz,1H); AB: 3.28 (d, J=7.5 Hz, 1H); 3.24 (d, J=7.5 Hz, 1H), 3.26 (d, J=6.1Hz, 2H); 2.96 (m, 1H); 2.88-2.75 (m, 2H); 2.71 (ddd, J=4.5, 9.0, 13.0Hz, 1H), 2.58 (ddd, J=7.0, 10.0, 14.0 Hz, 1H); 2.35 (m, 1H); 2.21 (m,1H); 2.00-1.80 (m, 6H); 1.40-1.20 (m, 1H); 1.17 (s, 3H); 0.93 (s, 3H);0.89 (dd, J=9.7 and 4.2 Hz, 1H). Mass spectrum (ESI) m/z (MH)⁺391.0.

[0215] Compound 40.

[0216] N-(2,2-Diphenylethyl)-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheylethane-1,2-diamine. 33 mg, 23% yield. ¹H NMR: δ7.31-7.18 (m, 10H), 4.13 (t, J=7.5 Hz, 1H), 3.27 (d, J=8.0 Hz, 2H), 3.14(dt, J=6.0, 10 Hz, 1H), (4H), 2.36 (qd, J=2.0, 6.0 Hz, 1H), 2.34 (dt,J=2.0, 10 Hz, 1H), 2.07-1.96 (m, 3H), 1.82 (dt, J=2.0, 6.0 Hz, 1H), 1.71(ddd, J=2.5, 5.5, 13.5 Hz, 1H), 1.22 (s, 3H), 1.09 (d, J=7.0 Hz, 3H),0.96 (d, J=10.5 Hz, 1H), 0.91 (s, 3H). Mass spectrum (ESI) m/z(MH)⁺377.3.

[0217] Compound 47.

[0218] N-(−)-cis-Myrtanyl-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheylethane-1,2-diamine. 42 mg, 33% yield. ¹H NMR: δ3.35-3.20 (m, 6H), 2.93 (dd, J=4.6, 2.0 Hz, 2H), 2.45-2.33 (m, 4H), 2.17(s, 3H), 2.06 (quint, J=7.0 Hz, 1H), 2.0-1.9 (m, 6H), 1.90 (dd, J=2.1,5.2 Hz, 1H), 1.87 (dt, J=1.8, 4.6 Hz, 1H), 1.51 (ddd, J=4.6, 10.0, 13.0Hz, 1H), 1.23 (s, 3H), 1.19 (s, 3H), 1.12 (d, J=8 Hz, 3H), 1.03 (d,J=10.3 Hz, 1H), 0.98 (s, 3H), 0.94 (d, J=9.8 Hz, 1H), 0.94 (s, 3H). Massspectrum (ESI) m/z (MH)⁺333.6.

[0219] Compound 52.

[0220] N-(3,3-Diphenylpropyl)-N′-cyclooetylethane-1,2-diamine. 20 mg,18% yield. ¹H NMR (500 MHz): δ 7.30-7.10 (m, 10H); 3.96 (t, J=7.9 Hz,1H); 3.00 (m, 1H); 2.90 (dd, J₁=J₂=5.5 Hz, 2H); 2.84 (dd, J₁=J₂=5.0 Hz,2H); 2.61 (t, J=7.3 Hz, 2H), 2.27 (q, J=7.6 Hz, 2H); 1.83 (m, 2H); 1.74(m, 2H); 1.65-1.40 (m, 10H).

[0221] Compound 55.

[0222] N-(1-Adamantylmethyl)-N′-cyclooctylethane-1,2-diamine. 6.7 mg, 6%yield. ¹H NMR (500 MHz): δ 3.08-3.02 (m, 1H), 3.02-2.98 (m, 2H);2.97-2.92 (m, 2H); 2.36 (s, 2H); 1.98 (m, 2H); 1.93-1.86 (m, 2H);1.80-1.50 (m, 19H).

[0223] Compound 57.

[0224] N-(−)-cis-Myrtanyl-N′-(cyclooctyl)ethane-1,2-diamine. 18 mg, 18%yield. ¹H NMR (500 MHz) δ 3.05-2.95 (m, 4H); AB: 2.76 (d, J=7.5 Hz, 1H),2.23 (d, J=8.0 Hz, 1H); 2.76 (dd, J=11.6 and 7.3 Hz, 1H); 2.73 (dd,J=11.9 and 8.2 Hz, 1H); 2.40-2.34 (m, 1H); 2.28 (quintet, J=8.0 Hz, 1H);1.97 (s, 3H); 2.00-1.84 (m, 6H); 1.80-1.70 (m, 2H); 1.68-1.38 (m, 11H);1.18 (s, 3H); 0.97 (s, 3H); 0.92 (d, J=9.8 Hz, 1H). Mass spectrum (ESI)m/z (MH)⁺307.5.

[0225] Compound 58.

[0226] N-(2-Adamantyl)-N′-cyclooctylethane-1,2-diamine. 25 mg, 23%yield. ¹H NMR: δ 3.06 (m, 1H), 3.00 (t, J=6.1 Hz, 2H), 2.93 (t, J=5, 5Hz, 2H), 2.83 (br s, 1H), 1.96 (s, 3H), 1.92-1.80 (m, 10H), 1.80-1.50(m, 20H). Mass spectrum (ESI) m/z (MH)⁺305.1.

[0227] Compound 59.

[0228] N-(Cyclooctyl)-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheylethane-1,2-diamine.

[0229] 15 mg, 14% yield. ¹H NMR (400 MHz): δ 3.47 (dt, J=6.0, 10.0 Hz,1H), 3.40-3.28 (m, 7H), 2.44 (tq, J=2.0, 10.0 Hz, 1H), 2.36 (dtd, J=2.0,6.0, 10.0 Hz, 1H), 2.09 (dq, J=2.0, 7.2 Hz, 1H), 2.00-1.90 (m, 3H),1.88-1.78 (m, 2H), 1.78-1.63 (m, 4H), 1.65-1.30 (m, 8H), 1.18 (d, J=6.0Hz, 3H), 1.16 (s, 3H), 1.17 (d, J=7.2 Hz, 1H), 0.90 (s, 3H). Massspectrum (ESI) m/z (MH)⁺307.4.

[0230] Compound 62.

[0231]N-(−)-cis-Myrtanyl-N′-(1S)-(1-ethylcyclohexane)ethane-1,2-diamine. 48mg, 46% yield. ¹H NMR (500 MHz): δ 3.06-3.00 (m, 1H); 2.98-2.95 (m, 2H);2.92-2.84 (m, 1H); 2.79 (dd, J=11.9 and 7.0 Hz, 1H); 2.75 (dd, J=11.9and 7.9 Hz, 1H); 2.73 (m, 1H); 2.39 (m, 1H); 2.28 (quintet, J=8.5 Hz,1H); 2.00-1.86 (m, 6H); 1.82-1.76 (m, 2H); 1.68 (m, 2H); 1.54-1.42 (m,2H); 1.32-1.10 (m, 6H); 1.19 (s, 3H); 1.13 (d, J=6.7 Hz, 3H); 1.07 (dd,J=12 and 3 Hz, 2H); 1.02 (dd, J=12 and 3 Hz, 2H); 0.98 (s, 3H); 0.93 (d,J=9.7 Hz, 1H). Mass spectrum (ESI) m/z (MH)⁺306.9.

[0232] Compound 65.

[0233]N-trans-(2-phenylcyclopropyl)-N′-(1-adamanthyl)ethane-1,2-diamine. 18mg, 16% yield. Mass spectrum (ESI) m/z (MH)⁺311.3.

[0234] Compound 66.

[0235] N-(3,3-Diphenylpropyl)-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheylethane-1,2-diamine. 2 mg, 2% yield. ¹H NMR (500MHz) δ 7.26 (m, 10H); 3.96 (t, J=7.6 Hz, 1H); 3.09 (m, 1H); 2.92 (m,1H); 2.84 (m, 2H); 2.62 (m, 2H); 2.35 (m, 4H); 1.97 (s, 3H); 1.82 (m,1H); 1.68 (m, 1H); 1.21 (s, 3H); 1.12 (d, J=7.3 Hz; 3H); 1.01 (m, 1H);0.92 (s, 3H). Mass spectrum (ESI) m/z (MH)⁺391.4.

[0236] Compound 73.

[0237] N-(2-Adamantyl)-N′-[2-(2-methoxyphenyl)ethyl]ethane-1,2-diamine.21 mg, 19% yield. ¹H NMR: δ 7.22 (dd, J=8.2, 7.3 Hz, 1H), 7.14 (d, J=7.3Hz, 1H), 6.89 (d, J=7.1, Hz, 1H), 6.87 (d, J=8.2, Hz, 1H), 3.81 (s, 3H),3.06 (t, J=7.1 Hz, 2H), 3.06 (m, 2H), 3.01 (m, 2H), 2.93 (t, J=7.1, 2H),1.95 (br s, 2H), 1.90-1.80 (m, 7H), 1.78-1.66 (m, 6H), 1.59 (d, J=2.5Hz, 2H). Mass spectrum (ESI) m/z (MH)⁺329.4.

[0238] Compound 78.

[0239] N-2-Adamantyl-N′-2,3-dihydro-1H-inden-2-yl-ethane-1,2-diamine.4.3 mg, 3% yield. ¹H NMR: δ 7.20 (dd, J=4.9, 8.5 Hz, 2H), 7.14 (dd,J=5.5, 2.1 Hz, 2H), 3.71 (quint, J=6.1 Hz, 2H), 3.19 (dd, J=5.8, 15.9Hz, 2H), 3.13 (br.s, 1H), 3.05 (m, 4H), 2.86 (dd, J=4.8, 15.8 Hz, 2H),2.08 (m, 2H), 2.00 (m, 6H), 1.96-1.88 (m, 4H), 1.88-1.80 (m, 3H), 1.74(m, 4H), 1.68-1.60 (m, 2H). Mass spectrum (ESI) m/z (MH)⁺303.4.

[0240] Compound 109.

[0241] N-Geranyl-N′-(2-adamanthyl)ethane-1,2-diamine. 27 mg, 24% yield.¹H NMR (400 MHz): δ 5.40 (t, J=7.2 Hz, 1H), 4.78 (br s, 2H), 3.64 (d,J=7.6 Hz, 2H), 3.34 (m, 2H), 2.07 (m, 2H), 2.08-1.95 (m, 4H), 1.95-1.85(m, 4H), 1.82 (m, 2H), 1.88-1.70 (m, 4H), 1.70-1.62 (m, 3H), 1.67 (s,3H), 1.56 (s, 3H), 1.50 (s, 3H). Mass spectrum (ESI) m/z (MH)⁺307.4.

[0242] Compound 111.

[0243] N-Geranyl-N′-(2-ethylpiperidine)ethane-1,2-diamine. 44 mg, 42%yield. ¹H NMR (500 MHz): δ 5.22 (t, J=6.1 Hz, 1H); 5.04 (m, 1H), 3.52(d, J=7.3 Hz, 2H); 3.05-2.85 (m, 4H); 2.66 (m, 1H); 2.44 (m, 2H); 2.08(m, 4H); 1.80-1.50 (m, 2H); 1.70(s, 3H); 1.65 (s, 3H); 1.58 (s, 3H);1.50-1.35 (m, 2H), 0.89 (t, J 7.3, 3H). Mass spectrum (ESI) m/z(MH)⁺293.4.

[0244] Compound 116.

[0245] N-Geranyl-N′-allyl-N′-(cyclopentyl)ethane-1,2-diamine. 45 mg, 42%yield. ¹H NMR: δ 5.86 (ddd, J=10.0, 16.1, 6.7 Hz, 1H), 5.28 (d, J=15.9Hz, 1H), 5.25 (d, J=8.7 Hz, 1H), 5.23 (t, J=7.3 Hz, 1H), 5.30 (m, 1H),3.59 (d, J=7.3 Hz, 2H), 3.28 (br d, J=6.4 Hz, 2H), 3.16 (quintet, J=8.2Hz, 1H), 3.02 (m, 2H), 2.95-2.86 (m, 2H), 1.88-1.80 (m, 4H), 1.70 (s,3H), 1.74-1.66 (m, 3H), 1.65 (s, 3H), 1.58 (s, 3H), 1.56-1.50 (2H),1.50-1.40 (m, 2H). Mass spectrum (ESI) m/z (MH)⁺305.3.

[0246] Compound 117.

[0247] N-Geranyl-N′-diphenylmethylethane-1,2-diamine. 24 mg, 20% yield.¹H NMR (500 MHz): δ 7.40 (d, J=7.2 Hz, 4H); 7.29 (t, J=7.3 Hz, 4H); 7.21(t, J=7.0 Hz, 2H); 5.15 (t, J=7.5, 1H); 5.01 (m, 1H); 4.89 (br s, 1H);3.42 (d, J=7.0 Hz, 2H); 3.00-2.78 2.93 (m, 4H); 2.20-2.00 2.17 (m, 4H);1.63 (s, 3H); 1.59 (s, 3H); 1.56 (s, 3H). Mass spectrum (ESI) m/z(MH)⁺363.3.

[0248] Compound 125.

[0249] N,N′-bis-(−)-cis-Myrtanylpropane-1,2-diamine. 82 mg, 70% yield.¹H NMR (500 MHz): δ 3.62 (m, 1H); 3.18 (dd, J=13.7 and 3.7 Hz, 1H); 3.05(dt, J=11.5 and 7.5 Hz, 1H); 3.06-2.92 (m, 2H); 2.86 (dt, J=12.2 and 7.3Hz, 1H); 2.40 (m, 4H); 2.06-1.84 (m, 10H); 1.56-1.46 (m, 2H); 1.37 and1.36 (two d, J=6.7 and J=7.0 Hz, 3H); 1.20 (s, 3H); 1.19 (m, 3H), 0.99and 0.98 (two s, 3H) Hz, H); 0.97 (s, 3H); 0.94 (two d, J=10.1 Hz, 2H).Mass spectrum (ESI) m/z (MH)⁺346.9.

[0250] Compound 151.

[0251] [2-(2-Methoxy)phenylethyl]-N′-(1R, 2R, 3R,5S)-(−)-isopinocampheyl-ethane-1,2-diamine. 67 mg, 60% yield. ¹H NMR(500 MHz): δ 7.23 (t, J=5.8 Hz, 1H); 7.13 (dd, J=5.8 and 1.8 Hz, 1H);6.88 (m, 2H); 3.81 (s, 3H); 3.13 (m, 1H); 3.1-3.0 (m, 3H); 3.01 (t,J=7.0 Hz, 2H); 2.89 (t, J=7.0 Hz, 2H); 2.42-2.35 (m, 2H); 2.00 (m, 3H);1.82 (dt, J=6.0 and 2.0 Hz, 1H); 1.72 (ddd, J=2.5, 5.5, 13.5 Hz, 1H);1.22 (s, 3H) 1.13 (d, J=7.3 Hz, 3H). 0.99 (d, J=10.1 Hz, 1H); 0.93 (s,3H). Mass spectrum (ESI) m/z (MH)⁺331.5.

[0252]N-2-(2-Methoxyphenyl)ethyl-N′-allyl-N′-cyclopentyl-ethane-1,2-diamine. 8mg, 7% yield. ¹H NMR: δ 7.26 (dd, J=7.3, 8.5, 1H), 7.18 (d, J=7.2 Hz,1H), 6.91 (m, 2H), 5.61 ddd, (J=6.7, 17.0, 9.4 Hz, 1H), 5.13 (d, J=15.3Hz, 1H), 5.10 (d, J=9.2 Hz, 1H), 3.83 (s, 3H), 3.13 (dd, J=7.0, 6.7 Hz,2H), 3.10 (d, J=6.7 Hz, 1H), 3.00 (d, J=7.3 Hz, 1H), 3.05-2.90 (m, 2H),2.97 (dd, J=8.2, 6.1 Hz, 2H), 2.75 (t, J=6.1 Hz, 2H), 1.73 (m, 2H), 1.62(m, 2H), 1.50 (m, 2H), 1.22 (m, 2H). Mass spectrum (ESI) m/z (MH)⁺311.4.

[0253]N²-(3-Phenylpropyl)-N¹-[2-(4-fluorophenyl)ethyl]-1-phenylethane-1,2-diamine.23 mg, 19% yield. ¹H NMR: δ 7.35 (d, J=7.6 Hz, 2H), 7.34 (quart, J=7.Hz, 1H), 7.26 (d, J=6.4 Hz, 3H), 7.23 (d, J=7.6 Hz, 2H), 7.17 (dd,J=7.3, 6.4 Hz, 1H), 7.12 (d, J=7.0 Hz, 2H), 3.21 (m, 1H), 3.03 (ddd,J=4.2, 8.0, 12.8 Hz, 4H), 2.86 (t, J=8.0 Hz, 2H), 2.85-2.79 (m, J=12.Hz, 2H), 2.74-2.64 (m, 4H), 2.61 (t, J=7.7 Hz, 2H), 1.96 (quint, J=7.6Hz, 2H). Mass spectrum (ESI) m/z (MH)⁺377.3.

EXAMPLE VII M. Tuberculosis Rv0341p Lucs Drug Response

[0254] Substituted ethylene diamines, as described herein, were testedon Mycobacterium tuberculosis using high-throughout screening assay withrecombinant mycobacterial containing promoter fusion of luciferase toRv0341 EMB-inducible promoter. This assay quickly and reliablyidentifies antimycobacterial activity in compound mixtures and/or inindividual compounds. In this assay, bioluminescence increases when themycobacteria is tested against an active compound, or an active compoundmixture. During this assay, a theoretical yield of 100% was assumed forevery unpurified substituted ethylene diamine, and the activity of eachsample was compared to commercially available ethambutol (99.0% purity).Results were reported in LCPS, and % Max. LCPS based on the activity ofEMB at. 3.1 μM.

[0255] The substituted ethylene diamines were analyzed according to thefollowing procedure. The diamines were dried in a speed vacuum to anapproximate concentration of 6.3 mmoles per well. Each diamine, ordiamine mixture, was then resuspended or dissolved in 200 μl of methanolfor a concentration of 31.5 mM diamine(s). The diamine(s) solution wasdiluted to a concentration of 200 μM in 7H9 broth medium (a 1:15.75dilution of the 31.5 mM stock, followed by a 1:10 dilution; eachdilution in 7H9 broth medium). Next, 50 μl of the diluted diamine(s)solution was added to the first well of a row of twelve in an opaque,96-well plate. The 7H9 broth medium, 25 μl, was added to each of theremaining wells (#2-12) in the row. The diamine(s) solution in “wellone” was serially diluted by transferring 25 μl from “well one” to “welltwo”, and repeating a 25 μl transfer from “well two” to “well three”,and so on, on through “well eleven”. In “well eleven”, the extra 25 μlof solution was discarded. “Well twelve” was used as a growth control toassess background activity of the reporter strain. The plate was thencovered and incubated at 37° C. for 24 hours. Immediately prior toanalysis, the following substrates were prepared: a buffer solutioncontaining 50 mM HEPES at pH 7.0 and 0.4% Triton X-100. Then, 0.25 ml ofIM DTT, and 14 μl of 10 mg/ml luciferin in DMSO were added to 5 ml ofthe buffer solution. This final solution (50 μl) was added to each ofthe twelve wells, immediately after the incubation period had run. Theluminescence from each well was measured 20 minutes after the luciferinsubstrate was added, using a TOPCOUNT® (Downers, Grove, Ill.) NXTluminometer (55/well).

[0256] FIGS. 6-8 show typical assay data for the luciferase reporterstrain containing an Rv0341 EMB-inducible promoter with serial dilutionof 12 wells (1 row) of a 96-well library plate. FIG. 10 shows the numberof substituted ethylene diamines with at least 10% luciferase activity,based on the activity of ethambutol at 3.1 μM.

[0257]FIG. 6 represents typical assay data in the luciferase reporterstrain containing an Rv0341 EMB-inducible promoter. The data representsvalues obtained from the HTS Luc assay for compound mixtures of one row(row D) in the 96-well library. Row D was subject to several serialdilutions. The effectiveness of the compound mixture in the assay wasmeasured by the intensity of luminescence, and compared to ethambutol(100% intensity, 99% purity) at 3.1 μM. Each curve in FIG. 6 representsone well, or ten compounds. Results are reported in percent maximumLuminescence Count per Second (% Max. LCPS). During the screening, atheoretical 100% chemical yield was assumed for every unpurifiedcompound. Concentrations are given for a single compound. Based on thisinitial screening, 300+compound mixtures showed anti-TB activity.

EXAMPLE VIII Representative MIC Experiment

[0258] The Minimum Inhibition Concentration (MIC) is the concentrationof the growth inhibitor, here a substituted ethylene diamine, at whichthere is no multiplication of seeded cells. A microdilution method wasused to determine the MIC of the substituted ethylene diamines, capableof inhibiting the growth of Mycobacterium tuberculosis in vitro. In arepresentative MIC experiment, bacteria, the H37Rv strain ofMycobacterium tuberculosis (M.tb), was cultivated in 7H9 medium to adensity of 0.2 OD (optical density) at 600 nm. The bacterial culture wasthen diluted 1:100 in 7H9 broth medium. Stock solutions of isoniazid andethambutol were each prepared at 32 μg/ml in 7H9 medium. A 3.2 mg/mlsolution of isonizid and ethambutol were each prepared in water. Thesolutions were then filtered, and diluted 1:100 in 7H9 medium. Eachdrug, purchased from Sigma, was “laboratory use only” grade. A 10 mMsolution of each substituted ethylene diamine was prepared in methanol.Next, 100 μl of the 7H9 medium was added to each well in a 96-well plate(rows (A through H) x columns (1 through 12)). To the first wells inrows C through H was added an additional 80 μl of the 7H9 medium. Theisoniazid solution, 100 μl, was added to well Al, and the ethambutolsolution, 100 μl, was added to well B1. Six substituted ethylenediamines, 20 μl each, were added to wells C1 through H1 (column 1),respectively. A serial dilution of each substituted ethylene diamine andthe isoniazid and ethambutol controls, was performed across each row.For example, a serial dilution across row C₁-C₁₂ was done by mixing andtransferring 100 μl of the previous well to the next consecutive well.In each well in “column 12,” 100 μl of the final dilution was discarded.Next, 100 μl of the diluted H37Rv strain of M.tb was added to each well.The 96-well plate was then covered and incubated at 37° C. for 10 days.The plate was read for bacterial growth, or non-growth, using aninverted plate reader. The MIC was determined to be the lowestconcentration of substituted ethylene diamine that inhibited visiblegrowth of the M.tb.

[0259] A representative plate layout, listing concentration in eachwell, is shown in Table 9. Table 10 lists MIC and LD50 data for selectedcompounds. The LD50 is the concentration of the substituted ethylenediamine at which 50% of the cells (H37Rv strain of M.tb) are killed.Table 11 lists MIC data for purified substituted ethylene diamines incomparison to ethambutol (EMB). FIG. 9 shows the number of substitutedethylene diamine compounds with MIC activity at various concentrationlevels. TABLE 9 Concentration in Each Well (μM) Based on Columns 1-12DRUG Isoniazid 58.25 29.13 14.56 7.28 3.64 1.82 0.91 0.45 0.23 0.11 0.060.03 Ethambutol 28.75 14.38 7.19 3.60 1.80 0.90 0.45 0.22 0.11 0.06 0.030.01 Subst. Ethylene 500 250 125 62.5 31.25 15.63 7.81 3.91 1.96 0.980.49 0.24 Diamine

[0260] TABLE 10 Selectivity Index for Selected Compounds MIC LD50 MICLD50 Cmpd (uM) (uM) MW (ug/ml) (ug/ml) SI 6 7.813 20 536 4.187768 10.722.559836 34 7.813 32 508 3.969004 16.256 4.095738 37 15.625 32 496 7.7515.872 2.048 47 15.625 25 452 7.0625 11.3 1.6 57 15.625 18 426 6.656257.668 1.152 59 15.625 32 426 6.65625 13.632 2.048 65 15.625 60 4306.71875 25.8 3.84 109 1.953 32 450 0.87885 14.4 16.38505 111 7.813 44412 3.218956 18.128 5.63164 151 7.813 41 450 3.51585 18.45 5.247664

[0261] The above procedure was also used to examine batched compounds(10 compounds per well). Synthesized batches of substituted ethylenediamines were dried in speed vacuum and then resuspended in DMSO orsterile water to a concentration of 2.5 mg/ml. TABLE 11 MIC Data forPurified Samples Plate set-up INH 58.25 29.125 14.56 7.28 3.64 1.82 0.910.45 0.23 EMB 28.75 14.375 7.1875 3.594 1.797 0.898 0.449 0.2245 0.1125CMPD 500 250 125 62.5 31.25 15.625 7.813 3.9063 1.953 Avg INH MIC (uM)Avg INH MIC (uM) 0.91 0.91 Avg EMB MIC (uM) Avg EMB MIC (uM) Avg EMB AvgEMB 7.1875 8.37 7.25 7.25 BACTEC Cmpd MIC (uM) (EMB: 2.5 UG/ML) 1 250250 125 125 2 250 250 250 250 3 31.25 62.5 15.6 15.6 4 125 62.5 62.562.5 5 >500 500 500 500 6 7.813* 7.813 3.9 3.9 7 15.625* 7.813 3.9 3.9 8125 125 31.25 31.25 10 7.813* 15.625 7.8 7.8 11 31.25 contaminated 3.93.9 13 31.25 31.25 15.6 15.6 15 14 15.625” 15.625 7.8 7.8 15 >500 >500250 500 17 62.5 62.5 15.6 15.6 21 15.625* 31.25 7.8 7.8 22 31.25 31.257.8 15.6 23 31.25 31.25 15.6 15.6 24 125 125 31.25 31.25 27 125 62.515.6 31.25 28 125 62.5 31.25 31.25 29 62.5 62.5 31.25 62.5 31 31.2561.25 15.6 15.6 32 15.625* 15.625 7.8 7.8 33 62.5 62.5 31.25 31.25 347.813* 7.813 3.9 3.9 35 62.5 62.5 15.6 31.25 36 31.25 62.5 15.6 15.6 3715.625* 15.625 3.9 7.8 1.25 38 7.813 7.813 3.9 7.8 40 15.625* 15.625 7.87.8 41 31.25 15.625 15.6 15.6 42 31.25 31.25 1.95 3.9 43 31.25 31.25 3.97.8 12.5 47 15.625* 15.625 1.95 7.8 5 51 31.25 250 31.25 31.25 5215.625* 15.625 3.9 3.9 53 31.25 31.25 31.25 31.25 54 31.25 31.25 15.631.25 55 15.625* 15.625 15.6 15.6 25 56 500 >500 500 500 57 15.625*7.813 7.8 7.8 58 15.625* 15.625 7.8 7.8 5 59 15.625* 31.25 15.6 15.612.5 61 62.5 62.5 31.25 31.25 62 15.625* 31.25 15.6 31.25 63 62.5 62.531.25 62.5 64 31.25 31.25 31.25 31.25 65 15.625* 31.25 31.25 31.25 6615.625* 15.625 7.8 7.8 68 500 500 500 500 71 62.5 62.5 31.25 31.25 7362.5 15.6 15.6 76 62.5 62.5 31.25 31.25 77 31.25 31.25 15.6 15.6 7815.625* 31.25 15.6 15.6 79 31.25 31.25 15.6 15.6 103 31.25 31.25 62.562.5 107 500 500 250 250 109 1.953* 1.953 1.95 1.95 0.63 111 7.813*7.813 7.8 7.8 5 116 15.625* 15.625 7.8 15.6 12.5 117 7.813* 15.625 7.87.8 118 31.25 62.5 31.25 no data 119 125 62.5 cont no data contam 12515.625* 15.625 cont no data 6.25 134 >500 >500 500 no data 151 15.625*7.813 cont no data 6.25 164 62.5 125 cont no data 165 62.5 62.5 15.615.6

EXAMPLE IX Secondary Screening and Evaluation of Substituted EthyleneDiamines Against Drug Resistant Patient Isolates

[0262] Secondary screening was performed on some of the substitutedethylene diamine compounds to examine their activity against threeclinically resistant MDR patient isolates. MDR Strain TN576 isclassified as a W1 strain (STP^(R), INH^(R), RIF^(R), EMB^(R), ETH^(R),KAN^(R), CAP^(R)) strain TN587 is classified as a W strain (STP^(R),INH^(R), RIF^(R), EMB^(R), KAN^(R)), and the third strain TN3086 isclassified as a W1 strain (STP^(R), INH^(R), RIF^(R), EMB^(R), KAN^(R)).Each MDR strain is highly resistant to ethambutol with MIC valuesexceeding 12.5-25 μM. The MICs for the following substituted ethylenediamines, MP 116, MP 117, RL 241, compounds #59 and #109, weredetermined for all three patient isolates.

[0263] The results from this study are shown in Tables 12-13. Table 14characterizes each MDR strain according to its resistance. TABLE 12Screening of Substituted Ethylene Diamines Against Drug ResistantPatient Isolates - (MIC values in ug/ml) WT 576 587 3806 EMB 3.12 (or11.1 uM) 12.5-25 12.5-25 12.5-25 MP 116 6.25 3.15 6.25 3.15 MP 117 6.253.15 3.15 3.15 RL 241  1.5 (or 3.34 uM) 1.5  1.5  1.5 

[0264] TABLE 13 Screening of Substituted Ethylene Diamines Against DrugResistant Patient Isolates (MIC values in ug/ml) WT 576 587 3806 EMB1.6-1.8 50 50 50 Cmpd #59 0.05 (or 0.13 uM) 0.1 0.05 0.05 Cmpd #109 0.10(or 0.18 uM) 0.2 0.2 0.1

[0265] TABLE 14 Drug Resistance of Each MDR Strain Strain STP STP 2 INH1INH2 Rif Emb Eth Kan Cip Cap Cyc  576 W1 R R R R R R R R S R S  587 W RR R R R R S R S S S 3806 W1 R R R R S R

EXAMPLE X In Vivo Animal Studies

[0266] Animal models were used in the final stages of the drug discoverycycle to assess the anti-microbial efficacy of some substitutedethylanediamine compounds in a representative system of human diseasestate. The in vivo testing approach involves the inoculation of four-sixweek old C57BL/6 mice via aerosol, containing approximately 200 colonyforming units of M. tuberculosis H37Rv.

[0267] A. CFU Lung Study

[0268] Mice aerosolized with M. tuberculosis H37Rv were examined for 10to 12 weeks following inoculation. Drugs (substituted ethylene diamines)were administered via the esophageal cannula (gavage) 7 days/week,starting at either 14 or 21 days post infection. Bacterial load in thelungs of five mice per group were determined at approximately one-weekintervals by viable colony counts. The drugs tested were directlycompared to the front line anti-tuberculosis drug isoniazid, and to thesecond line drug, ethambutol. Isoniazid and ethambutol were tested at 25mg/kg and 100 mg/kg, respectively. The substituted ethylene diamines,compound 37, compound 59 and compound 109, were each tested at 1 mg/kgand 10 mg/kg. FIGS. 17 to 19 represent data from three, independent CFULung studies. In each study, the number of colony forming units (CFU)that were recoverable and cultivatable, were determined during varioustime intervals (days).

[0269] B. Lesion Study

[0270] The ability of compound 59 and compound 109 to prevent thedevelopment of gross pathology due to bacterial burden was determined inconjunction with the CFU/Lung Study. The gross pathology was determinedby visible quantitation of lesions on the surface of the lungs.Quantitation by inspection is a good surrogate for CFU determination,and directly correlates to the bacterial burden, as determined by theactual colony forming units. The lesions are first visibly examined, andthen the lungs are processed and plated for CFU quantification. Thelesion study demonstrates the ability of the drug to prevent thedevelopment of the disease pathology. FIG. 20 represents data from alesion study. The corresponding CFU results are shown in FIG. 19.

[0271] C. Toxicity Study

[0272] Toxicity was assessed using a dose escalation study. This studywas performed with ten C57BL/6 mice per candidate. Every two days, themice were administered an increased concentration of the drug, andmonitored for detrimental effects. The administration scheme was 50,100, 200, 400, 600, 800 and 1000 mg/kg. The maximum limit of 1000 kg/mgwas based on the goal of dose escalation, and the solubility of thedrugs in the delivery vehicle. Compound 37 was toxic in mice at 100kg/mg. Compound 59 and compound 109 were tolerated in mice at 1000 mg/kgand 800 mg/kg, respectively.

[0273] It should be understood that the foregoing relates only topreferred embodiments of the present invention, and that numerousmodifications, or alterations, may be made therein without departingfrom the spirit and scope of the invention. The entire text of eachreference mentioned herein is hereby incorporated, in its entirety, byreference.

EXAMPLE XI In Vitro Toxicity and Selectivity Indexes for Hit Compounds

[0274] Twenty six compounds (including 37, 59 and 109) were tested in anin vitro model of toxicity using monkey kidney cells (Vero) and humancervical cancer cells (HeLa) using methods well known to those skilledin the art. The data from this toxicity testing and the MIC data wereused to calculate a selectivity index (SI), the ratio of IC50:MIC (Table15). Selectivity Indexes were ranging from 1.76 to 16.67. Compound 109has the best selectivity index. TABLE 15 In vitro data forrepresentative compounds. Compound MIC (μM) Vero IC50 (μM) SI (IC50:MIC)66 15.6 28 1.76 40 15.6 25 1.88 41 3.13 19 2.05 59 15.6 36 2.30 55 15.634 2.32 57 11.7 22 2.40 37 7.8 32 4.10 38 6.25 33 5.28 111 7.81 45 5.7673 12.5 81 6.48 58 12.5 82 6.56 78 15.6 130 8.33 109 1.56 26 16.67

EXAMPLE XII In Vivo Efficacy of Ethambutol Analogues

[0275] Compounds 58, 59, 73, 109, and 111 were selected for in vivoefficacy studies in a mouse model of TB. Compounds 58 and 59 share thesame cyclooctyl fragment in their molecules; compounds 58, 73, and 109share adamantly moiety, and 109 and 111—the geranyl fragment (FIG. 22).

[0276] In these studies, 8-week old inbred female mice C57BL/6 wereintravenously infected with M. tuberculosis. 3 weeks following infectiondrug treatment was initiated (detailed protocol is provided). The drugswere administered orally by gavage. Mice were sacrificed at threetimepoints (15, 30, and 45 days post infection), and CFUs in spleen andlungs were determined (FIGS. 23 and 24). These studies demonstrated thatcompound 109 had activity at doses 1 and 10 mg/kg equal to that ofethambutol at 100 mg/kg.

[0277] Materials and Methods

[0278] Mice. Female C57BL/6 mice of 8 weeks old were purchased fromCharles River (Raleigh, N.C.), housed in BSL-2 facility of BIOCAL, Inc.(Rockville, Md.), and were allowed to acclimate at least 4 days priorinfection.

[0279] Mycobacteria. An example of frozen and thawed of M. tuberculosisH37Rv Pasteur was added to 5 ml 7H10 broth medium, with 0.5% BSA and0.05% Tween 80, incubated 1 week at 37° C., and then 1 ml was added into25 ml medium (2-d passage during 2 weeks). Culture was washed twice andresuspended in PBS with 0.5% BSA and 0.05% Tween 80, aliquoted andfrozen at −80° C. To determined CFU of the culture aliquot was thawed,and 10-fold dilutions will be plated on agar 7H9 and CFU count will becalculated 20 days later.

[0280] Infection: Frozen sample of culture was thawed, and diluted forconcentration about 10⁶ CFU/ml. Mice were infected with M. tuberculosisH37Rv intravenously through lateral tail vein in corresponded dose in0.2 ml of PBS.

[0281] Antimicrobial Agents. INH, EMB, Ethambutol Analogues.

[0282] Protocol of drug treatment: Treatment of mice with compounds wasinitiated 20 days following infection. Compounds were dissolved in 10%ethanol in water and administered by gavage (0.2 ml per mouse). Therapywas given 5 days per week and continued for four or six weeks. Two, fourand six weeks following chemotherapy start mice (6 mice per group) weresacrificed, lungs and spleens were removed and homogenized in sterile in2 ml PBS with 0.05% Tween-80. Homogenates were plated in serialdilutions on 7H10 agar dishes, and incubated at 37° C. CFU counts werecalculated three weeks later.

[0283] Statistic analysis. To analyze results of CFUs in organs ANOVAtest was performed; the significance of the differences was estimated byStudent's test, p<0.05 was considered statistically significant.

[0284] Results

[0285] In vivo activities of new compounds. The activities of thesecompounds are presented in FIGS. 21-24. In the experiment presented inFIGS. 21 (spleen) and 22 (lung) mice were infected with 5×10⁵ CFU M.tuberculosis H37Rv and chemotherapy was started 20 days followinginfection. Mice were treated with INH (25 mg/kg), EMB (100 mg/kg),compounds 73 and 109 (both 1 mg/kg and 10 mg/kg). The results indicatethat in the spleen, compounds 73 and 109 have activities equal to thatof EMB at 100 mg/kg (FIG. 21). In spleen there are no statisticaldifferences between activities of these compounds at 1 mg/kg or 10mg/kg. In the lung, compound 109 at concentration 10 mg/kg after 4 and 6weeks was more effective than EMB at 100 mg/kg. In the lung,statistically sufficient difference was shown for compound 109 atconcentrations 1 mg/kg and 10 mg/kg (FIG. 22). INH was the most activedrug in both spleen and lung.

[0286] Compounds 73 and 109 were also tested in shorter model with usinghigher dose of infection (FIGS. 23 and 24). Mice were infected with5×10⁶ CFU M. tuberculosis H37Rv and chemotherapy was started 15 daysfollowing infection. Mice were treated with INH (25 mg/kg), EMB (100mg/kg), compounds 109 (0.1 mg/kg, 10 mg/kg, and 25 mg/kg), 58, 73 and111 (all 25 mg/kg). Mice were treated for 4 weeks. In both the spleenand lung, compound 109 at concentrations 10 mg/kg and 25 mg/kg hadactivity equal to that of EMB at 100 mg/kg, and at concentration 0.1mg/kg minimal but sufficient difference with untreated control appearedafter 4 weeks of therapy (FIGS. 23 and 24). Statistically sufficientdifference between compounds 73 (25 mg/kg) and 109 (25 mg/kg) wasdetected. In the lung significant difference between activities of thesecompounds was not detected. Compounds 58 and 111 are active in vivo inboth spleen and lung; however, compounds 73 and 109 are preferable. Theresults of these experiments indicate that compounds 73 and 109 in lowconcentration show activity equal that of EMB at 100 mg/kg, and in somecases compound 109 shows higher activity.

[0287] Testing of compounds 111 and 59 was performed in B6 mice infectedwith 5×10⁵ CFU M. tuberculosis H37Rv and beginning chemotherapy 20 daysfollowing infection (FIGS. 25 and 26). Both compounds showedanti-tuberculosis activity at concentration 10 mg/kg comparable to thatof EMB at 100 mg/kg.

[0288] In all experiments, INH showed higher activity than EMB and othercompounds decreasing load of bacteria in organs on 2-3 logs during 4-6weeks of chemotherapy; new compounds similar to EMB (100 mg/kg)decreased load of bacteria on 1.0-2.0 logs. Among studied compounds 73and 109 are the most preferable, because the highest capacity todecrease mycobacteria in organs and its parameters of toxicity andpharmacology kinetics.

EXAMPLE XIII In Vivo Toxicity

[0289] Preliminary dose acceleration studies in mice have indicated thatcompound 109 can be well tolerated at doses up to 800 mg/kg and compound59 up to 1000 mg/kg. Compound 37 was fatal at doses 100 mg/kg (ClifBarry, NIAID, unpublished results).

[0290] Compound 109 was mostly used in the form of dihydrochloride atfive different doses, and 37—solely as hydrochloride salt at two doses.

[0291] Mice were given a one-time dose of the compounds atconcentrations 100, 300 or 1000 mg/kg using the gavage method. Each doseof each compound consisted of one group of 3 mice. Monitoring of themice was done twice a day for the duration of the experiment. Micesurviving one week post-drug administration were sacrificed; criticalorgans were aseptically removed and observed for abnormalities andevidence of drug toxicity. The MTD (mg/kg) is the highest dose thatresults in no lethality/tissue abnormality.

[0292] Methods:

[0293] 1. Treatment of mice: C57BL/6 female mice (6-8 weeks in age) aregiven a onetime dose of the compound at concentrations 100, 300 or 1000mg/kg using the gavage method. The compounds are dissolved in theappropriate concentration of ethanol in distilled water and administeredin a volume of 0.2 ml per mouse.

[0294] 2. Observation of mice: Mice will be observed 4 and 6 hours postadministration, then twice daily for one week. Survival and body weightof mice will be closely monitored throughout the study.

[0295] 3. Assessment of drug toxicity: Mice exhibiting signs of anyabnormal appearance or behavior or those remaining in a group in whichother mice did not survive to day 7 will be sacrificed for assessment ofdrug toxicity. Critical organs will be aseptically removed and observed;tissues from the liver, heart, and kidneys are extracted and placed into10% formalin solution. These fixed tissues are sectioned and examinedfor abnormalities resulting from drug toxicity.

[0296] These studies indicate that the maximum tolerated dose for thecompound 109 is 600 mg/kg (Table 16). No visible changes in organs wereobserved. Dose 800 mg/kg was fatal: out of a group of 3 mice, twoanimals died within 3 days (Table 17). Compound 37 was well tolerated atdoses 100 and 300 mg/kg. No visible changes in organs were observed.Additional experiments to evaluate maximum tolerated dose and in vivoefficacy for the compound 37 are being conducted. TABLE 16 Determinationof a maximum tolerated dose for the compounds 109 and 37 in mice. 109 at109 at 109 at 109 at 37 at 100 mg/kg 300 mg/kg 600 mg/kg 1000 mg/kg 100mg/kg Day of Day of Day of Day of Day of Day Mice death Mice death Micedeath Mice death Mice death Apr. 8, 2003 1 3 3 3 3 2-4 h 1 Apr. 9, 20032 3 3 3 2 2 2 Apr. 10, 2003 3 3 3 3 2 2 Apr. 11, 2003 4 3 3 3 1 4 2 Apr.13, 2003 6 3 3 3 0 6 2 Apr. 14, 2003 7 3 3 3 — 2

[0297] TABLE 17 Determination of a maximum tolerated dose for thecompounds 109 and 37 in mice. 37 at 37 at 109 as HCl salt 109 as TFAsalt 100 mg/kg 300 mg/kg at 800 mg/kg at 800 mg/kg Day of Day of Day ofDay of Date Day Mice death Mice death Mice death Mice death Apr. 29,2003 1 3 3 3 1 Apr. 30, 2003 2 3 3 2/1 2 1 May 1, 2003 3 3 3 1/1 3 1 May2, 2003 4 3 3 1 1 5.03. 5 3 3 1 1 5.04 6 3 3 1 1 May 5, 2003 7 3 3 1 1

EXAMPLE XIV Pharmacokinetic Studies of the Compounds 37, 59, and 109

[0298] Initially, analytical methods for determination of the compoundshad been developed that allowed to carry out all the PK experiments, seeFIG. 29. Here is a brief description of the experiment: (1) plasmaspiked with tested compounds and 10 uL of Terfenadine or plasma samples(200 uL) added; (2) ACN (2 mL) added to precipitate protein and spin at2,500 rpm; (3) evaporate supernatant to dryness; (4) add 200 uL of thediluting solvent: methanol (with 0.1% of trifluoroacetic acid) ammoniumacetate (80/20); (5) vortex, spin, and use supernatant; (6) run LC/MS/MSon Sciex API 3000.

[0299] Biostability studies of the compounds in plasma were carried outusing concentrations 1 and 15 mg/ml. The compounds were incubated for 1,2, 3 & 6 hr at 37° C. (Table 18). In addition, it was found that alltested compounds were stable in plasma at 24° C., pH 2 and 7.4 up to 24hr. TABLE 18 Biostability of tested compounds in plasma. Comp. Human DogRat Mouse 37 20% ↓ stable 35% ↓ stable 25 59 stable stable stable stable109 30% ↓ 40% ↓ stable stable 30

[0300] Pilot PK study of the compounds 37, 59, and 109 in mice wasconducted using a cassette dosing: all the three analogs were formulatedtogether in saline at 1.5 mg/mL, and administered to mice simultaneouslyorally at 25 mg/kg, peritoneally at 6 mg/kg, and intravenously. It wasfound that doses 15 and 7.5 mg/kg caused death of mice, 3.75 mg/kgappeared lethargic immediately after dosing but then appeared normalappearance a few minutes later; 3 mg/kg displayed no adverse reactionsand hence was used as intravenous dose. Obtained data are presented onFIGS. 30, 31, and 32 (tested compounds were studied under the NCI'indexation NSC) and summarized in Table 19. TABLE 19 PK Parameters oftested compounds 37, 59, and 109 after a cassette dosing to mice. N/A -not detectable. Route i.v. i.v. i.v. i.p. i.p. i.p. p.o. p.o. p.o.Compounds 37 59 109 37 59 109 37 59 109 Dose (mg/kg) 3 3 3 6 6 6 25 2525 AUC (ng · h/mL) 954 384 1006 1372 272 1099 1602 169 655 Cmax (ng/mL)970 296 1192 630 217 935 263 28.7 227 T½ (h) 4.8 6.4 5.5 4.9 9.7 4.4 N/AN/A N/A CL (mL/kg/h) 3530 8043 3240 Bioavailabiiity (%) 72 35 55 3.3 0.92.7 Urine excretion (%) .71 1.9 .92 <0.01 <0.01 <0.01 N/A N/A N/A

[0301] Conducted pharmacokinetic studies indicated that compound 59 (NSC722040 by the NCI index) has relatively poor PK profiling (AUC, Cmax)and further testing of this compound was abandoned. Based on preliminarytoxicity data compound 37 was also ruled out as possible candidate.Therefore, compound 109 (NSC 722041 by the NCI) was selected for furtherPK analyses.

[0302] It has been shown that compound SQ109 reaches and exceeds itsMinimum Bactericidal Concentration MBC (313 ng/ml) in plasma whenadministered either iv or inatraveneously orally (p.o.), has a half-lifeof 5.2 h, and has total clearance less than hepatic blood flow (FIG. 33,Table 20). TABLE 20 Pharmacokinetic parameters of the compound 109.Parameters i.v. p.o. Dose (ma/ka) 3 25 AUC (ng · h/mL) 792 254 T_(1/2)ei (h) 3.5 5.2 C_(max) (ng/mL) 1038 135 T_(max) (h) 0 0.31 CL (mL/kg/h)3788 Vd_(SS) (mL/kg) 11826 Bioavailability 3.8

[0303] Its oral bioavailability is only 3.8% when administered p.o butthis is explained by its unique tissue distribution pattern. Tissuedistribution studies have demonstrated that SQ109 primarily distributesinto the lungs and spleen (FIGS. 34 and 35), which is highlyadvantageous for a infection that characteristically manifests as a lungdisease.

[0304] By using an ultracentrifugation method, it was found that plasmaprotein binding of the compound 109 is concentration dependent andvaries from 15% (20 ng/ml) to 74% (200 ng/ml) to 48% (2000 ng/ml). Afteri.v. dosing (3 mg/kg) the compound distributes between plasma and redblood cells in a ratio 70.6:29.4.

[0305] Little is known of the fate of the compound in the body, sincethe total amount of the compound after excretion (urine and feces) doesnot exceed 3% of the delivered dose (Table 2). TABLE 21 Amounts of thecompound 109 cumulatively excreted in mouse urine and feces followingsingle administration. Period after dosing (h) Dose/ Total Route Samples0-4 4-8 8-24 24-32 0-32  3 mg/kg Urine <0.01 <0.01 0.03 0.01 0.04 i.v.Feces <0.01 0.01 0.04 <0.01 0.06 25 mg/kg Urine — — — — p.o. Feces 0.480.31 1.12 0.08 2.0

[0306] Initial attempts to identify metabolites of the compound 109 inurine, did not provide evidence of breakdown products, FIG. 36. Forexample, there was no evidence for the formation of conjugatedmetabolites (M⁺521) in the mouse urine during first 24 hr followingcompound's administration, FIG. 37. Conjugated metabolites are productsof the typical metabolic pathway N-glucoronidation formed by reactionwith glucuronic acid (D. A. Williams and T. L. Lemke in Foye'sPrincipals of Medicinal Chemistry, 5^(th) Ed., p.202).

EXAMPLE XV In vitro Pharmacokinetic Studies of Compound 109

[0307] In vitro Pharmacology and early ADMET (Absorption, Distribution,Metabolism, Excretion, Toxicity) studies of the compound 109 werecontracted out to CEREP (15318 NE 95^(th) Street, Redmond, Wash. 98052,USA, www.cerep.com, tel 425 895 8666) under a Service Agreement andincluded testing against 30 standard receptors (see CEREP Tables 22 and23, provided in FIGS. 38 and 39, five CYP450 enzymes, hERG (K⁺ channel),aqueous solubility, predicted intestinal permeability, and metabolicstability (data presented in FIG. 40 Tables 24(a-m)).

EXAMPLE XVI Bis(2-Adamantyl)ethylenediamine, SQBisAd

[0308]

[0309] Compounds with the best Selectivity Indexes, such as 109, 58, 73,78, (Table 15) and good in vivo data share the same adamantane fragment(FIG. 20). A compound that would have solely this fragment (on bothsides of the ethylene linker) was contemplated. During preparation oftargeted 100,000 compound library of ethambutol analogues, 70,000compounds were proven to be formed, but 30,000 were failures. Thisparticular compound was not initially detected perhaps because it wassynthesized in very low yield or because it was never made due to stericfactors.

[0310] In the synthetic scheme used for preparation of the libraryScheme 1 (FIG. 41), sterically hindered amines on the second step rarelygave products. Analyzing MS data for a number of original plates it canbe stated that 2-adamantanamine when used as R₁NH₂ seldom yielddesirable products and this can be explained because of existence ofsterically hindered reaction site on the step 2 or step 3 of thesynthesis Scheme 2 (FIG. 41).

[0311] Compound SQBisAd can be prepared by “wet chemistry” using thesame route, Scheme 3 (FIG. 41), it is documented that 2-adamantamine(used as commercially available hydrochloride) does provide productswhen used on the 1 and 2 steps. Due to the symmetrical nature, thiscompound can be synthesized by alternative routes. We have preparedSQBisAd by reductive alkylation of ethylnediamine by 2-adamantanoneusing sodium cyanoborohydride. Final product (without additionalpurification) demonstrated MIC (Minimal Inhibitory Concentration) equalor better than compound 109.

EXAMPLE VIII Generating the Diamine Library with a Modified Linker

[0312] General Methods: All reagents were purchased from Sigma-Aldrich.Rink acid resin was purchased from NovaBiochem, Inc. Solventsacetonitrile, dichloromethane, dimethylformamide, ethylene dichloride,methanol, and tetrahydrofuran were purchased from Aldrich and used asreceived. Solid phase syntheses were performed on Quest 210 Synthesizer(Argonaut Technologies) and combinatorial chemistry equipment (WhatmanPolyfiltronics and Robbins Scientific). Evaporation of the solvents wasdone using SpeedVac AES (Savant). Mass spectra data were obtained byElectrospray Ionization technique on Perkin Elmer/Sciex, API-300, TQMSwith an autosampler.

[0313] The activation of the Rink-resin, the addition of the amine, andthe acylation step were carried out in 10 ml tubes using the Quest 210Synthesizer. Removal of the FMOC group, reductive alkylation reactionwith carbonyl compounds, the reduction with Red-A1, and the cleavagefrom the solid support were carried out in 96-deep (2 ml) well,chemically resistant plates.

[0314] Step 1. Activation of the Rink-Acid Resin.

[0315] A suspension of the Rink-acid resin (coverage of 0.43-0.63mmol/g), 6 g (up to 3.78 mmol), in 80 ml of 2:1 mixture ofdichloromethane and THF was disitrubuted into 20 tubes, 4 ml per tube,filtered and washed twice with THF. A solution of triphenylphosphine(5.7 g, 21.75 mmol) in 40 ml of THF was added, 2 ml/tube, followed bythe addition of a solution of hexachloroethane (5.09 g, 21.45 mmol) in20 ml of THF, 1 ml/tube. After 6 h the resins were washed with THF (2×4ml) and dichloromethane (2×4 ml).

[0316] Step 2. Addition of the First Amine.

[0317] Each tube was charged with 3 ml of dichloroethane, EtNiPr₂, (0.2ml, 1.15 mmol), and the corresponding amine (1 mmol). (When a selectedamine was a solid, it was added as a solution or a suspension in DMF).Dichloroethane was added to each tube to fill up the volume 4 ml. Thereaction was carried for 8 h at 45° C. and 6-8 h at room temperature.The resins were filtered, washed with a 2:1 mixture of dichloromethaneand methanol (1×4 ml), then with methanol (2×4 ml), and suck dry.

[0318] Step 3. Acylation with Fmoc Protected Amino Acid.

[0319] The resins were pre-washed with dichloromethane (2×4 ml). Eachtube was charged with 2 ml of dichloromethane, HATU (2 mol excess toloaded resin, 0.14 g, 0.39 mmol, dissolved in 1 ml of DMF), and 0.47mmol (2.5 mol excess to loaded resin) of amino acid dissolved in 1 ml ofDMF, and allowed to stir for 8 h at 45° C. and 6-8 h at roomtemperature. After 16 h the resins were filtered, washed with 1:1mixture of DMF and dichloromethane (1×3 ml), dichloromethane (1×3 ml)and acylation was repeated with the same amount of reagents. At the end,the resins were filtered, washed with 1:1 mixture of DMF anddichloromethane (1×3 ml), and methanol (3×3 ml), sucked dry (on Quest)for 30 min and transferred into vials (one resin per vial), and dried ina desiccator under vacuum for 1 h. After this step all resins weresubjected for quality control using MS spectra.

[0320] Step 4. Alkylation of the Amino Group.

[0321] Deprotection. Ten prepared resins from the first three steps werepooled together, leaving approximately 0.05 g of each in the individualvials for all necessary deconvolutions. A suspension of the resinmixture (2.0-2.5 g) in 100 ml of a 2:1 mixture of dichloromethane andTHF was distributed into two 96-well filterplates and filtered using afiltration manifold. The reaction plates were transferred intocombiclamps, and 0.2 ml of 20% solution of piperidine in DMF was addedto remove Fmoc protecting group and allowed to stay for 10 min. After 10min plate was filtered, washed with 0.2 ml of DMF, and deprotection wasrepeated with 0.2 ml of 20% solution of piperidine in DMF and allowed tostay for 20 min. After that plate was filtered, washed with DMF (0.2 mlper well) and dichloromethane (2×0.5 ml per well).

[0322] Reaction with the carbonyl compounds. Each well in row A on thereaction plate was charged with 0.1 ml of dichloromethane, 0.08 ml of˜1.0M solution of appropriate acid in DMF from master plate, 0.05 ml DMFsolution of PyBrop, (0.015 g, 0.03 mmol, 2.5 mol excess to loaded resin)and 0.05 ml of EtNiPr₂ in dichloromethane (0.022 ml, 0.13 mmol, 10 molexcess to loaded resin). Each well in rows B through H was charged with0.1 ml of THF, 0.160 ml of ˜1.0 M solution of appropriate aldehyde orketone in DMF from master plate and allowed to react for 30 min. After30 min 0.075 ml (0.075 mmol) of 1.0 M solution of NaBCNH₃ were added.The reaction plates were sealed and kept at RT for 72 h. At the end, theresins were filtered, washed with THF, DCM (1×1 ml), methanol (2×1 ml)and dried in desiccator under vacuum for 2 h.

[0323] Step 5. Reduction with Red-A1.

[0324] The reaction plates were placed into combiclamps. A 1:6 mixtureof Red-A1 (65+w % in toluene) and THF was added, 0.6 ml per well (0.28mmol of Red-A1 per well), and allowed to react for 4 h. After thereaction completion the resins were filtered, washed with THF (2×1 ml),methanol (3×1 ml) and dried in the filtration manifold.

[0325] Step 6. Cleavage.

[0326] This step was carried out using a cleavage manifold. The reactionplates (placed on the top of the collection plates in this manifold)were charged with a 10:85:5 mixture of TFA, dichloromethane, andmethanol, 0.5 ml per well. After 15 min, the solutions were filtered andcollected into proper wells of the collection plates. The procedure wasrepeated. Solvents were evaporated on a speedvac, and the residualsamples were ready for testing.

[0327] Deconvolution Example.

[0328] Deconvolution of the active wells was performed by re-synthesisof discrete compounds, from the archived FMOC-protected □-aminoacetamideresins (10 resins, 0.05-00.10 g each), which were set aside at the endof the acylation step before the pooling. Each resin was assigned adiscrete column (1, or 2, or 3, etc.) in a 96-well filterplate, and wasdivided between X rows (A, B, C, etc), where X is the number of hitsdiscovered in the original screening plate. To each well, in a row, aselected carbonyl compound (present in the hit) was added along withother required reagents: the first selected carbonyl compound was addedto the resins in the row “A”, the second carbonyl compound—to the resinsin the row “B”, the third carbonyl compound—to the resins in the row“C”, etc. A lay-out of a representative 96-well deconvolution plate isshown in Table 28, FIG. 52.

[0329] The reaction plates were sealed and kept at RT for 72 h. At theend, the resins were filtered, washed with THF, DCM (1×1 ml), methanol(2×1 ml) and dried in desiccator under vacuum for 2 h. Reduction andcleavage were performed according to steps 5 and 6 of the syntheticprotocol. The product wells from the cleavage were analyzed by ESI-MS(Electrospray Ionization Mass Spectroscopy) to ensure the identity ofthe actives, and were tested in the MIC assay. A summary of the ESI-MSdata is provided below. A list of compound hits and structures isprovided in Table 30, FIG. 53.

[0330] Compound 673

[0331]N²-[(2-methoxy-1-naphthyl)methyl]-3-phenyl-N¹-(3-phenylpropyl)propane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺439.2

[0332] Compound 674

[0333]N2-[2-(benzyloxy)ethyl]-N′-(3,3-diphenylpropyl)-4-(methylthio)butane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺463.4.

[0334] Compound 675

[0335]N¹-(3,3-diphenylpropyl)-4-(methylthio)-N²-(3-phenylpropyl)butane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺447.2

[0336] Compound 676

[0337]N²-(cyclohexylmethyl)-N¹-(3,3-diphenylpropyl)-4-(methylthio)butane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺425.1

[0338] Compound 677

[0339]N¹-(3,3-diphenylpropyl)-N²-(2-ethoxybenzyl)-4-(methylthio)butane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺463.1

[0340] Compound 678

[0341]N2-[2-(benzyloxy)ethyl]-N¹-[(6,6-dimethylbicyclo[3.1.1]hept-2-yl)methyl]-4-(methylthio)butane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺405.3

[0342] Compound 679

[0343]N¹-[(6,6-dimethylbicyclo[3.1.1]hept-2-yl)methyl]-4-(methylthio)-N-2-(3-phenylpropyl)butane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺389.5

[0344] Compound 680

[0345]N²-(2-chloro-4-fluorobenzyl)-4-methyl-N¹-(4-methylbenzyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺363.3, 365.5; (MCH₃CN) 403.3, 405.3.

[0346] Compound 681.

[0347]N²-[2-(benzyloxy)ethyl]-N¹-[2-(4-methoxyphenyl)ethyl]-4-methylpentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺385.1.

[0348] Compound 682.

[0349]N2-[3-(4-chlorophenoxy)benzyl]-N¹-[2-(4-methoxyphenyl)ethyl]-4-methylpentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺467.1, 469.2.

[0350] Compound 683.

[0351]N²-(4-isopropylbenzyl)-N¹-[2-(4-methoxyphenyl)ethyl]-4-methylpentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺383.3

[0352] Compound 684.

[0353]N¹-[2-(4-methoxyphenyl)ethyl]-4-methyl-N-2-[(2E)-3-phenylprop-2-enyl]pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺367.3; [M—(CH₂CH═CHPh)2H]+251.

[0354] Compound 685

[0355]N²-[2-(benzyloxy)ethyl]-4-methyl-N′-(3-phenylpropyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺369.1.

[0356] Compound 686.

[0357]N²-(2-chloro-4-fluorobenzyl)-4-methyl-N′-(3-phenylpropyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺377.2, 378.9.

[0358] Compound 687.

[0359]N2-[3-(4-chlorophenoxy)benzyl]-4-methyl-N¹-(3-phenylpropyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺451.1, 453.3.

[0360] Compound 688.

[0361]N²-(4-isopropylbenzyl)-4-methyl-N′-(3-phenylpropyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺367.3.

[0362] Compound 689

[0363]4-methyl-N²-[(2E)-3-phenylprop-2-enyl]-N′-(3-phenylpropyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺351.2.

[0364] Compound 690

[0365]N²-(2-ethoxybenzyl)-4-methyl-N¹-(3-phenylpropyl)pentane-1,2-diamine.Mass spectrum (ESI) m/z (MH)⁺369.1.

[0366] Compound 691.

[0367] N²-decahydronaphthalen-2-yl-N¹-[2(4-fluorophenyl)ethyl]-3-thien-3-ylpropane-1,2-diamine. Mass spectrum(ESI) m/z (MH)⁺415.3.

REFERENCES

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We claim:
 1. A composition comprising a substituted ethylene diaminecompound of the formula

wherein R₄ is selected from H, alkyl, aryl, heteroatom substituted alkyland aryl, alkenyl, alkynyl, aralkyl, aralkynyl, cycloalkyl,cycloalkenyl; and wherein R₁, R₂ and R₃ are independently selected fromH, alkyl, aryl, alkenyl, alkynyl, aralkyl, aralkenyl, aralkynyl,cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl, halide, alkoxy,aryloxy, alkylthio, arylthio, silyl, siloxy, amino; or wherein R₁ isselected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl, aralkenyl,aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl, halide,alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy, amino, and NR₂R₂ isderived from a cyclic secondary amine.
 2. The composition of claim 1,wherein NHR₁ or NR₂R₃ of the substituted ethylene diamine has thechemical structure


3. The composition of claim 2, wherein the substituted ethylene diaminecompound is selected from


4. The composition of claim 1, wherein NHR₁ or NR₂R₃ of the substitutedethylene diamine has the chemical structure


5. The composition of claim 4, wherein the substituted ethylene diaminecompound is selected from


6. The composition of claim 1, wherein NHR₁ or NR₂R₃ of the substitutedethylene diamine has the chemical structure


7. The composition of claim 6, wherein the substituted ethylene diaminecompound is selected from


8. The composition of claim 1, wherein NHR₁ or NR₂R₃ of the substitutedethylene diamine has the chemical structure


9. The composition of claim 8, wherein the substituted ethylene diaminecompound is selected from


10. The composition of claim 9, wherein the substituted ethylene diaminecompound is


11. The composition of claim 1, wherein the substituted ethylene diaminecompound is selected from


12. A method of preparing a substituted ethylene diamine compound of theformula

wherein R₄ is selected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl,aralkynyl, cycloalkyl, cycloalkenyl; and wherein R₁, R₂ and R₃ areindependently selected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl,aralkenyl, aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl,halide, alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy, amino; orwherein R₁ is selected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl,aralkenyl, aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl,halide, alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy, amino, andNR₂R₂ is derived from a cyclic secondary amine; comprising activating asolid-support resin containing hydroxyl groups with a halo-donatingreagent in the presence of base to produce a solid-support resincontaining halo groups; displacing the halo groups with an initialprimary amine to produce a solid-support resin containing amine groups;acylating the amine groups with a halo-acylhalide in the presence of abasic compound, or with a halo-acylacid in the presence of base, toproduce a solid-support resin containing α-haloacetyl amide groups;displacing α-halo groups of the α-haloacetyl amides with a secondary orsubsequent primary amine to produce a solid-support resin containinga-amine imide groups; reducing the carbonyl moiety on the a-amine imidegroups with a reducing agent to produce a solid-support resin containingtwo amine groups separated by two carbon atoms; cleaving the aminegroups separated by two carbon atoms from the solid support resin in thepresence of acid to produce the substituted ethylene diamine compound.13. The method of claim 12, wherein the initial primary amine is R₁NH₂.14. The method of claim 12, wherein the secondary or subsequent primaryamine is R₂R₃HN.
 15. A method of treating disease caused by aninfectious agent comprising administering an effective amount of asubstituted ethylene diamine compound of the formula:

wherein R₄ is selected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl,aralkynyl, cycloalkyl, cycloalkenyl; and wherein R₁, R₂ and R₃ areindependently selected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl,aralkenyl, aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl,halide, alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy, amino; orwherein R₁ is selected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl,aralkenyl, aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl,halide, alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy, amino, andNR₂R₂ is derived from a cyclic secondary amine.
 16. The method of claim15, wherein the infectious agent comprises a bacterial, mycological,parasitic, or viral agent.
 17. The method of claim 16, wherein thebacterial agent comprises M. tuberculosis, M. avium-intracellulare, M.kansarii, M. fortuitum, M. chelonae, M. leprae, M. africanum, M.microti, M. avium paratuberculosis, M. intracellulare, M. scrofulaceum,M. xenopi, M. marinum, or M. ulcerans.
 18. The method claim 15, whereinthe infectious disease comprises tuberculosis or Crohn's disease. 19.The method claim 15, wherein the substituted ethylene diamine compoundis


20. The method of claim 19, further comprising a pharmaceutical carrier.21. A composition comprising, a substituted ethylene diamine compoundcomprising,


22. A method of preparing a substituted ethylene diamine compound of theformula

wherein R₄ is selected from H, alkyl, aryl, heteroatom substituted alkyland aryl, alkenyl, alkynyl, aralkyl, aralkynyl, cycloalkyl,cycloalkenyl; and wherein R₁, R₂ and R₃ are independently selected fromH, alkyl, aryl, alkenyl, alkynyl, aralkyl, aralkenyl, aralkynyl,cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl, halide, alkoxy,aryloxy, alkylthio, arylthio, silyl, siloxy, amino; comprisingactivating a solid-support resin containing hydroxyl groups with ahalo-donating reagent in the presence of base to produce a solid-supportresin containing halo groups; displacing the halo groups with an initialprimary amine to produce a solid-support resin containing amine groups;acylating the amine groups with a FMOC protected amino acid in thepresence of a coupling reagent and a base, followed by removal of FMOCprotecting group to produce a solid-support resin containing α-aminoacetamide groups; modification of α-amino groups of the α-aminoacetamide groups with a carbonyl compound to produce a solid-supportresin containing corresponding derivative of α-amino acetamide groups;reducing the carbonyl moiety on the amide groups with a reducing agentto produce a solid-support resin containing two amine groups separatedby two carbon atoms; and cleaving the amine groups separated by twocarbon atoms from the solid support resin in the presence of acid toproduce the substituted ethylene diamine compound.
 23. A method fortreating an infectious disease comprising administering apharmaceutically effective amount of the composition in claim
 21. 24. Acomposition comprising a symmetrical substituted ethylene diaminecompound of the formula

wherein R₄ is selected from H, alkyl, aryl, heteroatom substituted alkyland aryl, alkenyl, alkynyl, aralkyl, aralkynyl, cycloalkyl,cycloalkenyl; and wherein R₁, R₂ and R₃ are independently selected fromH, alkyl, aryl, alkenyl, alkynyl, aralkyl, aralkenyl, aralkynyl,cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl, halide, alkoxy,aryloxy, alkylthio, arylthio, silyl, siloxy, amino; or wherein R₁ isselected from H, alkyl, aryl, alkenyl, alkynyl, aralkyl, aralkenyl,aralkynyl, cycloalkyl, cycloalkenyl, heteroalkyl, heteroaryl, halide,alkoxy, aryloxy, alkylthio, arylthio, silyl, siloxy, amino, and NR₂R₂ isderived from a cyclic secondary amine.